Cuantificación de TIMP-2 en plasmas de sangre de cordón umbilical de neonatos colombianos y su correlación con variables neonatales utilizando el método CBA en citometría de flujo
El Inhibidor tisular de las metaloproteinasas 2 (TIMP-2) cumple un papel esencial en el funcionamiento de la barrera hematoencefálica, la regulación de la neuroinflamación y renovación del hipocampo. Su concentración se encuentra aumentada en el plasma de sangre de cordón umbilical (SCU) y disminuye...
- Autores:
-
Cárdenas López, Valerie Niyireth
Guevara Sotelo, Laura Vanessa
- Tipo de recurso:
- Trabajo de grado de pregrado
- Fecha de publicación:
- 2019
- Institución:
- Colegio Mayor de Cundinamarca
- Repositorio:
- Repositorio Colegio Mayor de Cundinamarca
- Idioma:
- spa
- OAI Identifier:
- oai:repositorio.unicolmayor.edu.co:unicolmayor/214
- Acceso en línea:
- https://repositorio.unicolmayor.edu.co/handle/unicolmayor/214
- Palabra clave:
- Medicina regenerativa
Ingeniería de tejidos
Embriología
TIMP-2
CBA
Alzheimer
hipocampo
Plasma de sangre de cordón umbilical
- Rights
- openAccess
- License
- Derechos Reservados -Universidad Colegio Myor de Cundinamarca ,2019
id |
UCOLMAYOR2_e6ec6e7c8f9ffbe5fccbffc9c9ea9387 |
---|---|
oai_identifier_str |
oai:repositorio.unicolmayor.edu.co:unicolmayor/214 |
network_acronym_str |
UCOLMAYOR2 |
network_name_str |
Repositorio Colegio Mayor de Cundinamarca |
repository_id_str |
|
dc.title.spa.fl_str_mv |
Cuantificación de TIMP-2 en plasmas de sangre de cordón umbilical de neonatos colombianos y su correlación con variables neonatales utilizando el método CBA en citometría de flujo |
title |
Cuantificación de TIMP-2 en plasmas de sangre de cordón umbilical de neonatos colombianos y su correlación con variables neonatales utilizando el método CBA en citometría de flujo |
spellingShingle |
Cuantificación de TIMP-2 en plasmas de sangre de cordón umbilical de neonatos colombianos y su correlación con variables neonatales utilizando el método CBA en citometría de flujo Medicina regenerativa Ingeniería de tejidos Embriología TIMP-2 CBA Alzheimer hipocampo Plasma de sangre de cordón umbilical |
title_short |
Cuantificación de TIMP-2 en plasmas de sangre de cordón umbilical de neonatos colombianos y su correlación con variables neonatales utilizando el método CBA en citometría de flujo |
title_full |
Cuantificación de TIMP-2 en plasmas de sangre de cordón umbilical de neonatos colombianos y su correlación con variables neonatales utilizando el método CBA en citometría de flujo |
title_fullStr |
Cuantificación de TIMP-2 en plasmas de sangre de cordón umbilical de neonatos colombianos y su correlación con variables neonatales utilizando el método CBA en citometría de flujo |
title_full_unstemmed |
Cuantificación de TIMP-2 en plasmas de sangre de cordón umbilical de neonatos colombianos y su correlación con variables neonatales utilizando el método CBA en citometría de flujo |
title_sort |
Cuantificación de TIMP-2 en plasmas de sangre de cordón umbilical de neonatos colombianos y su correlación con variables neonatales utilizando el método CBA en citometría de flujo |
dc.creator.fl_str_mv |
Cárdenas López, Valerie Niyireth Guevara Sotelo, Laura Vanessa |
dc.contributor.advisor.none.fl_str_mv |
Ávila Portillo, Luz Mabel López Castro, Omaira Yaneth |
dc.contributor.author.none.fl_str_mv |
Cárdenas López, Valerie Niyireth Guevara Sotelo, Laura Vanessa |
dc.contributor.corporatename.spa.fl_str_mv |
Universidad Colegio Mayor de Cundinamarca |
dc.contributor.researchgroup.spa.fl_str_mv |
Trabajo de grado |
dc.subject.lemb.none.fl_str_mv |
Medicina regenerativa Ingeniería de tejidos Embriología |
topic |
Medicina regenerativa Ingeniería de tejidos Embriología TIMP-2 CBA Alzheimer hipocampo Plasma de sangre de cordón umbilical |
dc.subject.proposal.spa.fl_str_mv |
TIMP-2 CBA Alzheimer hipocampo Plasma de sangre de cordón umbilical |
description |
El Inhibidor tisular de las metaloproteinasas 2 (TIMP-2) cumple un papel esencial en el funcionamiento de la barrera hematoencefálica, la regulación de la neuroinflamación y renovación del hipocampo. Su concentración se encuentra aumentada en el plasma de sangre de cordón umbilical (SCU) y disminuye con el aumento de la edad, contribuyendo al deterioro progresivo del hipocampo. Estudios en modelo animal sugieren que el plasma de sangre de cordón umbilical contiene TIMP-2 y que los ratones viejos adquieren habilidades de memoria y temporoespaciales posterior a la infusión de plasmas jóvenes. Dado lo anterior, esta proteína está siendo investigada para el tratamiento de enfermedades neurodegenerativas como el Alzheimer. Este trabajo se realizó en STEM medicina Regenerativa, donde se obtuvo y procesó el plasma de SCU de 60 neonatos. Para la identificación y cuantificación de TIMP-2, se utilizó la técnica Cytometric Bead Array (CBA), con el Kit LEGENDplexTM Human TIMP Mix and Match. Los archivos FCS arrojados por el citómetro de flujo FACSCantoTM II fueron analizados por el software especializado LEGENDplex, y por medio de herramientas estadísticas como shapiro-wilk, coeficiente correlación de rango de Spearman y el programa estadístico STATA versión 12 del Hospital Militar Central de Bogotá, se realizó el análisis estadístico. Se encontro correlacion de TIMP-2 con el volumen, CD34+ y CNT. Sin embargo, no se encontro correlacion de TIMP-2 con las demás variables neonatales consideradas |
publishDate |
2019 |
dc.date.issued.none.fl_str_mv |
2019-11 |
dc.date.accessioned.none.fl_str_mv |
2021-06-10T20:34:55Z |
dc.date.available.none.fl_str_mv |
2021-06-10T20:34:55Z |
dc.type.spa.fl_str_mv |
Trabajo de grado - Pregrado |
dc.type.coar.spa.fl_str_mv |
http://purl.org/coar/resource_type/c_7a1f |
dc.type.coarversion.spa.fl_str_mv |
http://purl.org/coar/version/c_970fb48d4fbd8a85 |
dc.type.content.spa.fl_str_mv |
Text |
dc.type.driver.spa.fl_str_mv |
info:eu-repo/semantics/bachelorThesis |
dc.type.redcol.spa.fl_str_mv |
https://purl.org/redcol/resource_type/TP |
dc.type.version.spa.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
http://purl.org/coar/resource_type/c_7a1f |
status_str |
publishedVersion |
dc.identifier.uri.none.fl_str_mv |
https://repositorio.unicolmayor.edu.co/handle/unicolmayor/214 |
dc.identifier.barcode.none.fl_str_mv |
60144 |
url |
https://repositorio.unicolmayor.edu.co/handle/unicolmayor/214 |
identifier_str_mv |
60144 |
dc.language.iso.spa.fl_str_mv |
spa |
language |
spa |
dc.relation.ispartof.none.fl_str_mv |
No objeto asociado |
dc.relation.references.spa.fl_str_mv |
1. Harper, E. Collagenases. Annual Review of Biochemistry. [Internet] 1980. Vol 49(1): 1063–1078. [Cited 12 Apr 2019]. Disponible en: https://www.annualreviews.org/doi/abs/10.1146/annurev.bi.49.070180.005215?journalCode=biochem 2. Stetler-Stevenson W, Krutzsch H, Liotta L. Tissue inhibitor of metalloproteinase (TIMP-2). A new member of the metalloproteinase inhibitor family. The Journal of Biological Chemistry. [Internet] 1989. Vol 264 (29): 17374-17378. [Cited 18 sep 2018]. Disponible en: http://www.jbc.org/content/264/29/17374.long 3. Rosenberg GA, Kornfeld M, Estrada E, Kelley RO, Liotta L A & Stetler-Stevenson WG. TIMP-2 reduces proteolytic opening of blood-brain barrier by type IV collagenase. Brain Research. [Internet]1992. Vol 576 (2): 203-207. [Cited 18 sep 2018]. Disponible en: https://www.ncbi.nlm.nih.gov/pubmed/1381261 4. Blavier L, DeClerck A. Tissue inhibitor of metalloproteinases-2 is expressed in the interstitial matrix in adult mouse organs and during embryonic development. Mol Biol Cell. [Internet] 1997. Vol 8(8): 1513–1527. [Cited 22 sep 2018]. Disponible en: https://www.ncbi.nlm.nih.gov/pubmed/9285822 5. Shields, L. E., & Andrews, R. G. Gestational age changes in circulating CD34+ hematopoietic stem/progenitor cells in fetal cord blood. American Journal of Obstetrics and Gynecology. [Internet] 1998. 178(5), 931–937. [Cited 28 sep 2019]. Disponible en: https://www.ncbi.nlm.nih.gov/pubmed/9609562 6. BIOSCIENCES, B. D. Introduction to Flow Cytometry: A learning guide. Manual Part. [Internet] 2000; vol. 1, no 1. [Cited 22 Apr 2019]. Disponible: https://www.bu.edu/flow-cytometry/files/2010/10/BD-Flow-Cytom-Learning-Guide.pdf 7. Janowska-Wieczorek, A. Differential MMP and TIMP production by human marrow and peripheral blood CD34+ cells in response to chemokines. Experimental Hematology. [Internet] 2000; 28(11), 1274–1285. [Cited 16 Sep 2019]. Disponible en: https://www.ncbi.nlm.nih.gov/pubmed/11063876 8. Ballen, K., Wilson, M., Wuu, J., Ceredona, A., Hsieh, C., Stewart, F., … Quesenberry, P. Bigger is better: maternal and neonatal predictors of hematopoietic potential of umbilical cord blood units. Bone Marrow Transplantation. [Internet] 2001; 27(1):7-14. [Cited 16 Sep 2019]. Disponible en: https://www.ncbi.nlm.nih.gov/pubmed/11244432 9. Maymon, E., Romero, R., Pacora, P., Gomez, R., Mazor, M., Edwin, S., … Berry, S. M. A role for the 72 kDa gelatinase (MMP-2) and its inhibitor (TIMP-2) in human parturition, premature rupture of membranes and intraamniotic infection. Journal of Perinatal Medicine, [Internet] 2001; 29(4). [Cited 22 Sep 2019]. Disponible en: https://www.ncbi.nlm.nih.gov/pubmed/11565199 10. Morgan, E., Varro, R., Sepulveda, H., Ember, J. A., Apgar, J., Wilson, J., … Gaur, A. Cytometric bead array: a multiplexed assay platform with applications in various areas of biology. Clinical Immunology. [Internet] 2004; Vol 110(3): 252–266. [Cited 29 apr 2019]. Disponible en: https://www.sciencedirect.com/science/article/pii/S1521661603003139 11. Nakagawa, R., Watanabe, T., Kawano, Y., Kanai, S., Suzuya, H., … Kaneko, M. Analysis of maternal and neonatal factors that influence the nucleated and CD34+ cell yield for cord blood banking. TRANSPLANTATION AND CELLULAR ENGINEERING. [Internet] 2004; 44(2). [Cited 29 sep 2019]. Disponible en: https://www.ncbi.nlm.nih.gov/pubmed/14962318 12. Jaworski D, Boone J, Caterina J, Soloway P, Falls W. Prepulse inhibition and fear-potentiated startle are altered in tissue inhibitor of metalloproteinase-2 (TIMP- 54 2) knockout mice. Brain Research. [Internet] 2005; 1051: 81-89. [Cited 22 sep 2018]. Disponible en: https://www.ncbi.nlm.nih.gov/pubmed/15979591 13. Jaworski D, Soloway P, Caterina J, Falls W. Tissue Inhibitor of Metalloproteinase-2 (TIMP-2) deficient mice display motor deficits. J Neurobiol. [Internet] 2006. Vol 66(1): 82–94. [Cited 25 sep 2018]. Disponible en: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1440718/ 14. Cockle, J. V., Gopichandran, N., Walker, J. J., Levene, M. I., & Orsi, N. M. Matrix Metalloproteinases and Their Tissue Inhibitors in Preterm Perinatal Complications. Reproductive Sciences, [Internet] 2007; 14(7), 629–645. [Cited 22 Sep 2019]. Disponible en: https://www.ncbi.nlm.nih.gov/pubmed/18000225 15. Montalvo, A. (dir). Evaluación genética y microanalítica de las células madre de de la gelatina de Wharton para su utilización en ingeniería tisular. [tesis doctoral en Internet]. [España]: Universidad de Granada; 2008; 242 p. [citado 26 de Abr de 2019]. Disponible en: https://hera.ugr.es/tesisugr/1768058x.pdf 16. DANE, Ministerio de Salud y Protección Social. Glosario. Estadísticas Vitales – Módulo de Nacimientos y Defunciones del RUAF. [Internet]. Abril 2009. [Cited 7 may 2019]. Disponible en: https://www.minsalud.gov.co/Documentos %20y%20Publicaciones/Glosario%20EEVV.pdf 17. Brew K, Nagase H.The tissue inhibitors of metalloproteinases (TIMPs): An ancient family with structural and functional diversity [Review]. Biochimica et Biophysica Acta (BBA) - Molecular Cell Research. [Internet] 2010. Vol 1803 (1): 55-71. Disponible en: https://www.sciencedirect.com/science/article/pii/S0167488910000042 18. Ruiz C, Nariño D, Muñoz J. Epidemiología y carga de la Enfermedad de Alzheimer. Acta Neurol Colomb. [Internet] 2010; 26:Sup (3:1):87-94). [Cited 25 sep 2018]. Disponible en: https://www.acnweb.org/acta/acta_2010_26_Supl3_1_87-94.pdf 19. Madsen-Bouterse S, Romero R, Tarca A, Kusanovic J, Espinoza J, Kim C et al. The Transcriptome of the Fetal Inflammatory Response Syndrome. Am J Reprod Immunol. [Internet]. 2010 January; 63(1): 73–92. [Cited 26 sep 2018]. Disponible en: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3437779/ 20. Chandra T, Afree S, Kumar A, ingh U. Correlation of umbilical cord blood volume with CD34+ cells concentration. International Journal of Blood Transfusion and Immunohematology 2010; 1: 11-15. [Cited 26 sep 2018]. Disponible en: http://www.ijbti.com/archive/2011-archive/100003IJBTITC2011-chandra/100003IJBTITC2011-chandra-full-text.php 21. Wen S, Zhao W, Lin P, Yang K. cTransfusion and Apheresis Science [Internet] .2012; 46: 39–45 [Cited 26 sep 2018]. Disponible en: https://www.trasci.com/article/S1473-0502(11)00221-7/fulltext 22. Becton, Dickinson and Company. BD Cytometric Bead Array: Multiplexed Bead-Based Immunoassays. BD Biosciences. [Internet]. 2012. [Cited 29 apr 2019]. Disponible en: https://www.bdbiosciences.com/documents/CBA_Brochure_Intl.pdf 23. Chandra, T., Afreen, S., Kumar, A., Singh, U., y Gupta, A. Does Umbilical Cord Blood-derived CD34+ Cell Concentration Depend on the Weight and Sex of a Full-term Infant?. Journal of Pediatric Hematology / Oncology. [Internet]. 2012; 34(3):184-7. [Cited 16 sep 2019]. Disponible en: https://www.ncbi.nlm.nih.gov/pubmed/22441709 24. Wan, Z., You, S., Rong, Y., Zhu, B., Zhang, A., Zang, H., … Xin, S. CD34+ Hematopoietic Stem Cells Mobilization, Paralleled with Multiple Cytokines Elevated in Patients with HBV-Related Acute-on-Chronic Liver Failure. Digestive Diseases and Sciences, [Internet]. 2012; 58(2), 448–457. [Cited 20 sep 2019]. Disponible en: https://www.ncbi.nlm.nih.gov/pubmed/23095991 25. Kim, D., Staples, M., Shinozuka, K., Pantcheva, P., Kang, S.-D., & Borlongan, C. Wharton’s Jelly-Derived Mesenchymal Stem Cells: Phenotypic Characterization and Optimizing Their Therapeutic Potential for Clinical Applications. International Journal of Molecular Sciences. [Internet]. 2013; 14(6), 11692–11712. [Cited 05 May 2019]. Disponible en: https://www.mdpi.com/1422-0067/14/6/11692 26. Katsimpardi L, Litterman NK, Schein PA, Miller CM, Loffredo FS, Wojtkiewicz GR, et al. Vascular and neurogenic rejuvenation of the aging mouse brain by young systemic factors. Science. [Internet] 2014;344:630–634 [Cited 29 sep 2018]. Disponible en: https://www.ncbi.nlm.nih.gov/pubmed/24797482 27. Lee EJ, Kim HS. The anti-inflammatory role of tissue inhibitor of metalloproteinase-2 in lipopolysaccharide-stimulated microglia. J Neuroinflammation. [Internet] 2014; 11: 116 [Cited 29 sep 2018]. Disponible en: https://www.ncbi.nlm.nih.gov/pubmed/2497034 28. Bequer L, Gómez T, Pérez de Alejo L, Molina O, Peláez M, Ferrer M. Estudio bioquímico en sangre del cordón umbilical de recién nacidos teniendo en cuenta edad gestacional y peso al nacer. Revista del Laboratorio clínico. [Internet] 2014. Vol 7(3) 111-118 [Cited 01 oct 2018]. Disponible en: http://www.elsevier.es/es-revista-revista-del-laboratorio-clinico-282-articulo-estudio-bioquimico-sangre-del-cordon-S1888400814000610 29. Moodley KK., Chan D. The Hippocampus in Neurodegenerative Disease. The Hippocampus in Clinical Neuroscience. [Internet] 2014; 95–108 [Cited 01 oct 2018]. Disponible en: https://www.ncbi.nlm.nih.gov/pubmed/24777134 30. Al-Deghaither S. Impact of maternal and neonatal factors on parameters of hematopoietic potential in umbilical cord blood. Saudi Med J. [Internet] 2015 Jun; 36(6): 704–712 [Cited 01 oct 2018]. Disponible en: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4454905 31. Rodríguez F, Riaño D, León A, Mosquera A, Rojas A, Augusto C, et al. Correlación entre características físicas neonatales y maternas con el recuento total de células nucleadas y de células CD34+ por microlitro en sangre de cordón umbilical. Revista Med. [Internet] 2015; 23 (2): 71-77 [Cited 01 oct 2018]. Disponible en: https://revistas.unimilitar.edu.co/index.php/rmed/article/view/1749 32. Aguirre, E. La función del hipocampo en el procesamiento de la memoria y su deterioro durante el envejecimiento. Revista Mexicana de Neurociencia. [Internet] 2015; 16(4): 21-30 [Cited 02 oct 2018]. Disponible en: http://www.medigraphic.com/cgi-bin/new/resumen.cgi?IDARTICULO=64941 33. Castellano JM, Kirby ED, Wyss-Coray T.Blood-borne revitalization of the aged brain.JAMA Neurol. [Internet] 2015;72:1191–1194 [Cited 02 oct 2018]. Disponible en: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4867550/ 34. Garcia B, Rubio F, Crespo M. Tecnicas de analisis hematologico. Paraninfo. 2015. España: 258-259. 35. Olaya, M; Vargas, W; Bernal, J. Una aproximación desde la física a las consecuencias patológicas de la longitud excesiva del cordón umbilical. Revista Colombiana de Obstetricia y Ginecología. [Internet]. 2015; Vol. 66 No. 1 (53-60) [Cited 5 feb 2019]; . Disponible en: http://www.scielo.org.co/pdf/rcog/v66n1/v66n1a07.pdf 36. Olivares, J., Juárez, E., García, F. El hipocampo: neurogénesis y aprendizaje. Rev Med UV, [Internet] 2015; 20-28. [Cited 06 May 2019]. Disponible en: https://www.researchgate.net/profile/Fabio_Garcia-Garcia/publication/282251666_El_hipocampo_neurogenesis_y_aprendizaje/links/5609813a08ae840a08d3afa6.pdf 37. Lee, C., An, J., Kim, J. H., Kim, E. S., Kim, S. H., Cho, Y. K., … Sheen, Y. H. Low levels of tissue inhibitor of metalloproteinase-2 at birth may be associated with subsequent development of bronchopulmonary dysplasia in preterm infants. Korean Journal of Pediatrics, [Internet] 2015; 58(11), 415. [Cited 20 Sep 2019]. Disponible en: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4675921/ 38. Adan, A., Alizada, G., Kiraz, Y., Baran, Y., & Nalbant, A. Flow cytometry: basic principles and applications. Critical Reviews in Biotechnology [Internet] 2016; Vol 37(2), 163–176. [Cited 10 Apr 2019]. Disponible en: https://pdfs.semanticscholar.org/7e79/b4057a7a9f0cdaadca7c2dbb15e0e3c1b3cd.pdf 39. Castellano J, Mosher K, Abbey RJ, McBride A, James M, Berdnik D, Shen J, Zou B, Xie X, Tingle M, Hinkson I, Angst M, Wyss-Coray T. Human umbilical cord plasma proteins revitalize hippocampal function in aged mice. Nature. [Internet] 2017; Apr 27;544(7651):488-492. [Cited 10 oct 2018]. Disponible en: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5586222 40. Wang, L., Wang, D. Liu, M. Yao, R. Correlation between Tissue Characterization and Dynamic Expression of Matrix Metalloproteinase-2 and Its Tissue Inhibitor in Conjunctival Filtering Bleb of Rats. Biomed research international. [Internet] 2017; 2017, 10 pages. [Cited 16 Ene 2019]. Disponible en: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5624151/pdf/BMRI2017-1054129.pdf 41. Gayatri, R., Crasta, J., Thomas, T., Pratibha, D., Thomas, A., Sridhar, T. S., Kurpad, A. V. Structural Analysis of the Umbilical Cord and Its Vessels in Intrauterine Growth Restriction and Pre-eclampsia. Journal of Fetal Medicine. [Internet]. 2017; 4(2), 85–92. [Cited 27 Abr 2019]. Disponible en: https://link.springer.com/article/10.1007/s40556-017-0118-2 42. Mottet, N., Chaussy, Y., Arbez-Gindre, F., & Riethmuller, D. Fisiología y patologías del cordón umbilical. EMC - Ginecología-Obstetricia. [Internet]. 2017; 53(4), 1–12. [Cited 17 May 2019]. Disponible en: https://www.sciencedirect.com/science/article/abs/pii/S1283081X17868879 43. Sundrani D, Narang A, Mehendale S, Joshi S, Chavan-Gautam P. Investigating the expression of MMPs and TIMPs in preterm placenta and role of CpG methylation in regulating MMP-9 expression. IUBMB Journals. [Internet]. 2017; 69, 985-993 [Cited 18 Jul 2019]. Disponible en: https://iubmb.onlinelibrary.wiley.com/doi/full/10.1002/iub.1687 44. Zarkesh M, Zadeh-Vakili A, Azizi F, Fanaei SA, Foroughi F, Hedayati M. The Association of BRAF V600E Mutation With Tissue Inhibitor of Metalloproteinase-3 Expression and Clinicopathological Features in Papillary Thyroid Cancer. International Journal of Endocrinology and Metabolism. [Internet] 2018; 16(2): e56120. [Cited 16 oct 2018]. Disponible en: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5972213/ 45. Mashayekhi F, Saberi A, Mashayekhi S. Serum TIMP1 and TIMP2 concentration in patients with different grades of meningioma. Clinical Neurology and Neurosurgery. [Internet] 2018. 170, 84-87. [Cited 10 feb 2019]. Disponible en: https://www.sciencedirect.com/science/article/pii/S0303846718301756?via%3Dihub 46. Liu Z, Ren Y, Zhu F. Expression of MMP-2 and TIMP‑3 with incidence and prognosis of giant-cell tumor of the bone. Oncology Letters. Oncol Lett. [Internet] 2018; 16 (1): 721–726. [Cited 16 oct 2018]. Disponible en: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6019976/ 47. R&D Systems. Quantikine ELISA - Human TIMP-2 Immunoassay. Catálogo DTM200 [Internet]. 2018. [Cited 18 mar 2019]. Disponible en: https://resources.rndsystems.com/pdfs/datasheets/dtm200.pdf 48. Wang, W. Zhang, Y. Liu, M. Wang, Y. Yang, T. Li, D. Li, Q. TIMP2 is a Poor Prognostic Factor and Predicts Metastatic Biological Behavior in Gastric Cancer. Scientific Reports (Nature Publisher Group). [Internet]. 2018; 8, 1-11. [Cited 18 mar 2019]. Disponible en: https://ezproxy.unicolmayor.edu.co:2136/central/docview/2059010336/E4BDA59C6A134AE0PQ/1?accountid=50438 49. R&D SYSTEMS a biotechne brand. Quantikine ELISA Human TIMP-2 Immunoassay [Internet]. USA R&D Systems, Inc; 2018. [Cited 16 mar 2019]. Disponible en: https://resources.rndsystems.com/pdfs/datasheets/dtm200.pdf 50. Camacho, M., Escalona, R., Romero, S., Vargas, M., Zúñiga, M., Mora. J. Células madre de sangre de cordón umbilical: obtención, aplicaciones y situación en Costa Rica. REVISTA MÉDICA DE LA UNIVERSIDAD DE COSTA RICA. [Internet]. 2018; Vol 12 Nº 1. . [Cited 16 sep 2019]. Disponible en: https://revistas.ucr.ac.cr/index.php/medica/article/view/34612 51. Schaduangrat, N, Prachayasittikul, V, Choomwattana, S, Wongchitrat, P, Phopin, K, Suwanjang, W, et al. Multidisciplinary approaches for targeting the secretase protein family as a therapeutic route for Alzheimer’s disease. Medicinal Research Reviews. [Internet] 2019. [Cited 16 Mar 2019]. Disponible en: https://www.ncbi.nlm.nih.gov/pubmed/30628099 52. Medeiros, N. I., & Gomes, J. Cytometric Bead Array (CBA) for Measuring Cytokine Levels in Chagas Disease Patients. T. Cruzi Infection. [Internet] 2019; vol 1955 309–314. [Cited 29 Apr 2019]. Disponible en: https://link.springer.com/protocol/10.1007/978-1-4939-9148-8_23 53. Malemud CJ. Inhibition of MMPs and ADAM/ADAMTS. Biochem Pharmacol. [Internet] 2019;165:33-40. [Cited 29 sep 2019]. Disponible en: https://www.ncbi.nlm.nih.gov/m/pubmed/30826330/ 54. Biolegend® Enabling Legendary Discovery™. Human TIMP Mix and Match Subpanel. [Internet] Vol 2. [Cited 29 sep 2018]. Disponible en: https://www.biolegend.com/Files/Images/media_assets/pro_detail/datasheets/75084_Hu_TIMP_Mix_and_Match_Subpanel_V2.pdf |
dc.rights.eng.fl_str_mv |
Derechos Reservados -Universidad Colegio Myor de Cundinamarca ,2019 |
dc.rights.uri.spa.fl_str_mv |
https://creativecommons.org/licenses/by-nc-sa/4.0/ |
dc.rights.accessrights.spa.fl_str_mv |
info:eu-repo/semantics/openAccess |
dc.rights.creativecommons.spa.fl_str_mv |
Atribución-NoComercial-CompartirIgual 4.0 Internacional (CC BY-NC-SA 4.0) |
dc.rights.coar.spa.fl_str_mv |
http://purl.org/coar/access_right/c_abf2 |
rights_invalid_str_mv |
Derechos Reservados -Universidad Colegio Myor de Cundinamarca ,2019 https://creativecommons.org/licenses/by-nc-sa/4.0/ Atribución-NoComercial-CompartirIgual 4.0 Internacional (CC BY-NC-SA 4.0) http://purl.org/coar/access_right/c_abf2 |
eu_rights_str_mv |
openAccess |
dc.format.extent.spa.fl_str_mv |
70p. |
dc.format.mimetype.spa.fl_str_mv |
application/pdf |
dc.coverage.city.none.fl_str_mv |
Bogotá D.C. |
dc.publisher.spa.fl_str_mv |
Universidad Colegio Mayor de Cundinamarca |
dc.publisher.faculty.spa.fl_str_mv |
Facultad de Ciencias Sociales |
dc.publisher.place.spa.fl_str_mv |
Bogotá, Distrito Capital |
dc.publisher.program.spa.fl_str_mv |
Bacteriología y Laboratorio Clínico |
institution |
Colegio Mayor de Cundinamarca |
bitstream.url.fl_str_mv |
https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/214/1/Sustentaci%c3%b3n%20-%20TIMP-2.pdf https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/214/2/1%20PARA%20SUBIR%20Tesis%20-%20TIMP-2.pdf https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/214/3/2%20PARA%20SUBIR%20Tesis%20-%20TIMP-2%20%281%29.pdf https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/214/4/license.txt https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/214/5/Sustentaci%c3%b3n%20-%20TIMP-2.pdf.txt https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/214/7/1%20PARA%20SUBIR%20Tesis%20-%20TIMP-2.pdf.txt https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/214/9/2%20PARA%20SUBIR%20Tesis%20-%20TIMP-2%20%281%29.pdf.txt https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/214/6/Sustentaci%c3%b3n%20-%20TIMP-2.pdf.jpg https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/214/8/1%20PARA%20SUBIR%20Tesis%20-%20TIMP-2.pdf.jpg https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/214/10/2%20PARA%20SUBIR%20Tesis%20-%20TIMP-2%20%281%29.pdf.jpg |
bitstream.checksum.fl_str_mv |
d5149307b9ac41f611913f07cea31ae0 157f1da9550ed17e43c700a4cc45ef38 ca4206b2f79670f656fe0deb8d080854 2f9959eaf5b71fae44bbf9ec84150c7a c8753b61349839bd0633274e6fa698af d00c31b5323af4579136df57a96e9af9 e1c06d85ae7b8b032bef47e42e4c08f9 c0ff83d31576e4ff81eb95c8eddc6d45 5fb0bd793d9aacad85f82745fe98a231 8f62c0a3866786294521b43bb5c58a0c |
bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 MD5 MD5 MD5 MD5 MD5 MD5 MD5 MD5 |
repository.name.fl_str_mv |
Biblioteca Digital Unicolmayor |
repository.mail.fl_str_mv |
repositorio@unicolmayor.edu.co |
_version_ |
1812210084959223808 |
spelling |
Ávila Portillo, Luz Mabele1163c132ab850431fd649d0078f064eLópez Castro, Omaira Yaneth11d3416123ec6bebf56c78bff186d9a5Cárdenas López, Valerie Niyireth2978c6e17f6bff0168106db6367661feGuevara Sotelo, Laura Vanessa65b8c0b25b20355f2a77bc14b10ed388Universidad Colegio Mayor de CundinamarcaTrabajo de grado2021-06-10T20:34:55Z2021-06-10T20:34:55Z2019-11https://repositorio.unicolmayor.edu.co/handle/unicolmayor/21460144El Inhibidor tisular de las metaloproteinasas 2 (TIMP-2) cumple un papel esencial en el funcionamiento de la barrera hematoencefálica, la regulación de la neuroinflamación y renovación del hipocampo. Su concentración se encuentra aumentada en el plasma de sangre de cordón umbilical (SCU) y disminuye con el aumento de la edad, contribuyendo al deterioro progresivo del hipocampo. Estudios en modelo animal sugieren que el plasma de sangre de cordón umbilical contiene TIMP-2 y que los ratones viejos adquieren habilidades de memoria y temporoespaciales posterior a la infusión de plasmas jóvenes. Dado lo anterior, esta proteína está siendo investigada para el tratamiento de enfermedades neurodegenerativas como el Alzheimer. Este trabajo se realizó en STEM medicina Regenerativa, donde se obtuvo y procesó el plasma de SCU de 60 neonatos. Para la identificación y cuantificación de TIMP-2, se utilizó la técnica Cytometric Bead Array (CBA), con el Kit LEGENDplexTM Human TIMP Mix and Match. Los archivos FCS arrojados por el citómetro de flujo FACSCantoTM II fueron analizados por el software especializado LEGENDplex, y por medio de herramientas estadísticas como shapiro-wilk, coeficiente correlación de rango de Spearman y el programa estadístico STATA versión 12 del Hospital Militar Central de Bogotá, se realizó el análisis estadístico. Se encontro correlacion de TIMP-2 con el volumen, CD34+ y CNT. Sin embargo, no se encontro correlacion de TIMP-2 con las demás variables neonatales consideradasResumen 8 1. Introducción 9 2. Objetivos 10 2.1. Objetivo general 10 2.2. Objetivos específicos 10 3. Antecedentes 11 4. Marco teórico 22 4.1. Fisiología y anatomía del Cordón umbilical 22 4.2. Sangre y plasma de sangre de Cordón Umbilical 23 4.3. Generalidades y funciones de los inhibidores de las metaloproteinasas (TIMP) 24 4.4 TIMP-2 25 4.4.1. Estudios en modelo animal. 26 4.4.2. Estudios en humanos 27 4.5. Técnicas utilizadas para la cuantificación de TIMP-2 28 4.5.1 ELISA (Método enzimático). 28 4.5.2. Citometría de Flujo. 28 4.5.3. Cytometric bead array CBA (perlas de captura citométricas). 30 4.6. Fisiología y anatomía del Hipocampo 32 4.7. Enfermedad de Alzheimer 32 5. Materiales y métodos 34 5.1. Clasificación del diseño de investigación 34 5.2. Población 34 5.3. Selección y tamaño de la muestra 34 5.4. Criterios de inclusión y de exclusión 34 Criterios de inclusión 34 Criterios de exclusión. 34 5.5. Variables 35 5.5.1. Variables neonatales y perinatales. 35 5.5.2. Variables dependientes. 35 5.5.3. Variables independientes. 35 5.6. Obtención y procesamiento de muestras 37 5.7. Determinación de TIMP-2. 37 6. Plan de análisis 39 6.1 Análisis de datos 39 6.2 Análisis estadístico 39 7. Resultados 40 8. Discusión 45 9. Conclusiones 49 10. Aspectos éticos 50 Referencias 52 Anexos 62 1. Consentimiento informado 62 2. Base de datos selección de muestras 66PregradoBacteriólogo(a) y Laboratorista ClínicoTrabajo de grado70p.application/pdfspaUniversidad Colegio Mayor de CundinamarcaFacultad de Ciencias SocialesBogotá, Distrito CapitalBacteriología y Laboratorio ClínicoNo objeto asociado1. Harper, E. Collagenases. Annual Review of Biochemistry. [Internet] 1980. Vol 49(1): 1063–1078. [Cited 12 Apr 2019]. Disponible en: https://www.annualreviews.org/doi/abs/10.1146/annurev.bi.49.070180.005215?journalCode=biochem2. Stetler-Stevenson W, Krutzsch H, Liotta L. Tissue inhibitor of metalloproteinase (TIMP-2). A new member of the metalloproteinase inhibitor family. The Journal of Biological Chemistry. [Internet] 1989. Vol 264 (29): 17374-17378. [Cited 18 sep 2018]. Disponible en: http://www.jbc.org/content/264/29/17374.long3. Rosenberg GA, Kornfeld M, Estrada E, Kelley RO, Liotta L A & Stetler-Stevenson WG. TIMP-2 reduces proteolytic opening of blood-brain barrier by type IV collagenase. Brain Research. [Internet]1992. Vol 576 (2): 203-207. [Cited 18 sep 2018]. Disponible en: https://www.ncbi.nlm.nih.gov/pubmed/13812614. Blavier L, DeClerck A. Tissue inhibitor of metalloproteinases-2 is expressed in the interstitial matrix in adult mouse organs and during embryonic development. Mol Biol Cell. [Internet] 1997. Vol 8(8): 1513–1527. [Cited 22 sep 2018]. Disponible en: https://www.ncbi.nlm.nih.gov/pubmed/92858225. Shields, L. E., & Andrews, R. G. Gestational age changes in circulating CD34+ hematopoietic stem/progenitor cells in fetal cord blood. American Journal of Obstetrics and Gynecology. [Internet] 1998. 178(5), 931–937. [Cited 28 sep 2019]. Disponible en: https://www.ncbi.nlm.nih.gov/pubmed/96095626. BIOSCIENCES, B. D. Introduction to Flow Cytometry: A learning guide. Manual Part. [Internet] 2000; vol. 1, no 1. [Cited 22 Apr 2019]. Disponible: https://www.bu.edu/flow-cytometry/files/2010/10/BD-Flow-Cytom-Learning-Guide.pdf7. Janowska-Wieczorek, A. Differential MMP and TIMP production by human marrow and peripheral blood CD34+ cells in response to chemokines. Experimental Hematology. [Internet] 2000; 28(11), 1274–1285. [Cited 16 Sep 2019]. Disponible en: https://www.ncbi.nlm.nih.gov/pubmed/110638768. Ballen, K., Wilson, M., Wuu, J., Ceredona, A., Hsieh, C., Stewart, F., … Quesenberry, P. Bigger is better: maternal and neonatal predictors of hematopoietic potential of umbilical cord blood units. Bone Marrow Transplantation. [Internet] 2001; 27(1):7-14. [Cited 16 Sep 2019]. Disponible en: https://www.ncbi.nlm.nih.gov/pubmed/112444329. Maymon, E., Romero, R., Pacora, P., Gomez, R., Mazor, M., Edwin, S., … Berry, S. M. A role for the 72 kDa gelatinase (MMP-2) and its inhibitor (TIMP-2) in human parturition, premature rupture of membranes and intraamniotic infection. Journal of Perinatal Medicine, [Internet] 2001; 29(4). [Cited 22 Sep 2019]. Disponible en: https://www.ncbi.nlm.nih.gov/pubmed/1156519910. Morgan, E., Varro, R., Sepulveda, H., Ember, J. A., Apgar, J., Wilson, J., … Gaur, A. Cytometric bead array: a multiplexed assay platform with applications in various areas of biology. Clinical Immunology. [Internet] 2004; Vol 110(3): 252–266. [Cited 29 apr 2019]. Disponible en: https://www.sciencedirect.com/science/article/pii/S152166160300313911. Nakagawa, R., Watanabe, T., Kawano, Y., Kanai, S., Suzuya, H., … Kaneko, M. Analysis of maternal and neonatal factors that influence the nucleated and CD34+ cell yield for cord blood banking. TRANSPLANTATION AND CELLULAR ENGINEERING. [Internet] 2004; 44(2). [Cited 29 sep 2019]. Disponible en: https://www.ncbi.nlm.nih.gov/pubmed/1496231812. Jaworski D, Boone J, Caterina J, Soloway P, Falls W. Prepulse inhibition and fear-potentiated startle are altered in tissue inhibitor of metalloproteinase-2 (TIMP- 54 2) knockout mice. Brain Research. [Internet] 2005; 1051: 81-89. [Cited 22 sep 2018]. Disponible en: https://www.ncbi.nlm.nih.gov/pubmed/1597959113. Jaworski D, Soloway P, Caterina J, Falls W. Tissue Inhibitor of Metalloproteinase-2 (TIMP-2) deficient mice display motor deficits. J Neurobiol. [Internet] 2006. Vol 66(1): 82–94. [Cited 25 sep 2018]. Disponible en: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1440718/14. Cockle, J. V., Gopichandran, N., Walker, J. J., Levene, M. I., & Orsi, N. M. Matrix Metalloproteinases and Their Tissue Inhibitors in Preterm Perinatal Complications. Reproductive Sciences, [Internet] 2007; 14(7), 629–645. [Cited 22 Sep 2019]. Disponible en: https://www.ncbi.nlm.nih.gov/pubmed/1800022515. Montalvo, A. (dir). Evaluación genética y microanalítica de las células madre de de la gelatina de Wharton para su utilización en ingeniería tisular. [tesis doctoral en Internet]. [España]: Universidad de Granada; 2008; 242 p. [citado 26 de Abr de 2019]. Disponible en: https://hera.ugr.es/tesisugr/1768058x.pdf16. DANE, Ministerio de Salud y Protección Social. Glosario. Estadísticas Vitales – Módulo de Nacimientos y Defunciones del RUAF. [Internet]. Abril 2009. [Cited 7 may 2019]. Disponible en: https://www.minsalud.gov.co/Documentos %20y%20Publicaciones/Glosario%20EEVV.pdf17. Brew K, Nagase H.The tissue inhibitors of metalloproteinases (TIMPs): An ancient family with structural and functional diversity [Review]. Biochimica et Biophysica Acta (BBA) - Molecular Cell Research. [Internet] 2010. Vol 1803 (1): 55-71. Disponible en: https://www.sciencedirect.com/science/article/pii/S016748891000004218. Ruiz C, Nariño D, Muñoz J. Epidemiología y carga de la Enfermedad de Alzheimer. Acta Neurol Colomb. [Internet] 2010; 26:Sup (3:1):87-94). [Cited 25 sep 2018]. Disponible en: https://www.acnweb.org/acta/acta_2010_26_Supl3_1_87-94.pdf19. Madsen-Bouterse S, Romero R, Tarca A, Kusanovic J, Espinoza J, Kim C et al. The Transcriptome of the Fetal Inflammatory Response Syndrome. Am J Reprod Immunol. [Internet]. 2010 January; 63(1): 73–92. [Cited 26 sep 2018]. Disponible en: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3437779/20. Chandra T, Afree S, Kumar A, ingh U. Correlation of umbilical cord blood volume with CD34+ cells concentration. International Journal of Blood Transfusion and Immunohematology 2010; 1: 11-15. [Cited 26 sep 2018]. Disponible en: http://www.ijbti.com/archive/2011-archive/100003IJBTITC2011-chandra/100003IJBTITC2011-chandra-full-text.php21. Wen S, Zhao W, Lin P, Yang K. cTransfusion and Apheresis Science [Internet] .2012; 46: 39–45 [Cited 26 sep 2018]. Disponible en: https://www.trasci.com/article/S1473-0502(11)00221-7/fulltext22. Becton, Dickinson and Company. BD Cytometric Bead Array: Multiplexed Bead-Based Immunoassays. BD Biosciences. [Internet]. 2012. [Cited 29 apr 2019]. Disponible en: https://www.bdbiosciences.com/documents/CBA_Brochure_Intl.pdf23. Chandra, T., Afreen, S., Kumar, A., Singh, U., y Gupta, A. Does Umbilical Cord Blood-derived CD34+ Cell Concentration Depend on the Weight and Sex of a Full-term Infant?. Journal of Pediatric Hematology / Oncology. [Internet]. 2012; 34(3):184-7. [Cited 16 sep 2019]. Disponible en: https://www.ncbi.nlm.nih.gov/pubmed/2244170924. Wan, Z., You, S., Rong, Y., Zhu, B., Zhang, A., Zang, H., … Xin, S. CD34+ Hematopoietic Stem Cells Mobilization, Paralleled with Multiple Cytokines Elevated in Patients with HBV-Related Acute-on-Chronic Liver Failure. Digestive Diseases and Sciences, [Internet]. 2012; 58(2), 448–457. [Cited 20 sep 2019]. Disponible en: https://www.ncbi.nlm.nih.gov/pubmed/2309599125. Kim, D., Staples, M., Shinozuka, K., Pantcheva, P., Kang, S.-D., & Borlongan, C. Wharton’s Jelly-Derived Mesenchymal Stem Cells: Phenotypic Characterization and Optimizing Their Therapeutic Potential for Clinical Applications. International Journal of Molecular Sciences. [Internet]. 2013; 14(6), 11692–11712. [Cited 05 May 2019]. Disponible en: https://www.mdpi.com/1422-0067/14/6/1169226. Katsimpardi L, Litterman NK, Schein PA, Miller CM, Loffredo FS, Wojtkiewicz GR, et al. Vascular and neurogenic rejuvenation of the aging mouse brain by young systemic factors. Science. [Internet] 2014;344:630–634 [Cited 29 sep 2018]. Disponible en: https://www.ncbi.nlm.nih.gov/pubmed/2479748227. Lee EJ, Kim HS. The anti-inflammatory role of tissue inhibitor of metalloproteinase-2 in lipopolysaccharide-stimulated microglia. J Neuroinflammation. [Internet] 2014; 11: 116 [Cited 29 sep 2018]. Disponible en: https://www.ncbi.nlm.nih.gov/pubmed/249703428. Bequer L, Gómez T, Pérez de Alejo L, Molina O, Peláez M, Ferrer M. Estudio bioquímico en sangre del cordón umbilical de recién nacidos teniendo en cuenta edad gestacional y peso al nacer. Revista del Laboratorio clínico. [Internet] 2014. Vol 7(3) 111-118 [Cited 01 oct 2018]. Disponible en: http://www.elsevier.es/es-revista-revista-del-laboratorio-clinico-282-articulo-estudio-bioquimico-sangre-del-cordon-S188840081400061029. Moodley KK., Chan D. The Hippocampus in Neurodegenerative Disease. The Hippocampus in Clinical Neuroscience. [Internet] 2014; 95–108 [Cited 01 oct 2018]. Disponible en: https://www.ncbi.nlm.nih.gov/pubmed/2477713430. Al-Deghaither S. Impact of maternal and neonatal factors on parameters of hematopoietic potential in umbilical cord blood. Saudi Med J. [Internet] 2015 Jun; 36(6): 704–712 [Cited 01 oct 2018]. Disponible en: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC445490531. Rodríguez F, Riaño D, León A, Mosquera A, Rojas A, Augusto C, et al. Correlación entre características físicas neonatales y maternas con el recuento total de células nucleadas y de células CD34+ por microlitro en sangre de cordón umbilical. Revista Med. [Internet] 2015; 23 (2): 71-77 [Cited 01 oct 2018]. Disponible en: https://revistas.unimilitar.edu.co/index.php/rmed/article/view/174932. Aguirre, E. La función del hipocampo en el procesamiento de la memoria y su deterioro durante el envejecimiento. Revista Mexicana de Neurociencia. [Internet] 2015; 16(4): 21-30 [Cited 02 oct 2018]. Disponible en: http://www.medigraphic.com/cgi-bin/new/resumen.cgi?IDARTICULO=6494133. Castellano JM, Kirby ED, Wyss-Coray T.Blood-borne revitalization of the aged brain.JAMA Neurol. [Internet] 2015;72:1191–1194 [Cited 02 oct 2018]. Disponible en: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4867550/34. Garcia B, Rubio F, Crespo M. Tecnicas de analisis hematologico. Paraninfo. 2015. España: 258-259.35. Olaya, M; Vargas, W; Bernal, J. Una aproximación desde la física a las consecuencias patológicas de la longitud excesiva del cordón umbilical. Revista Colombiana de Obstetricia y Ginecología. [Internet]. 2015; Vol. 66 No. 1 (53-60) [Cited 5 feb 2019]; . Disponible en: http://www.scielo.org.co/pdf/rcog/v66n1/v66n1a07.pdf36. Olivares, J., Juárez, E., García, F. El hipocampo: neurogénesis y aprendizaje. Rev Med UV, [Internet] 2015; 20-28. [Cited 06 May 2019]. Disponible en: https://www.researchgate.net/profile/Fabio_Garcia-Garcia/publication/282251666_El_hipocampo_neurogenesis_y_aprendizaje/links/5609813a08ae840a08d3afa6.pdf37. Lee, C., An, J., Kim, J. H., Kim, E. S., Kim, S. H., Cho, Y. K., … Sheen, Y. H. Low levels of tissue inhibitor of metalloproteinase-2 at birth may be associated with subsequent development of bronchopulmonary dysplasia in preterm infants. Korean Journal of Pediatrics, [Internet] 2015; 58(11), 415. [Cited 20 Sep 2019]. Disponible en: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4675921/38. Adan, A., Alizada, G., Kiraz, Y., Baran, Y., & Nalbant, A. Flow cytometry: basic principles and applications. Critical Reviews in Biotechnology [Internet] 2016; Vol 37(2), 163–176. [Cited 10 Apr 2019]. Disponible en: https://pdfs.semanticscholar.org/7e79/b4057a7a9f0cdaadca7c2dbb15e0e3c1b3cd.pdf39. Castellano J, Mosher K, Abbey RJ, McBride A, James M, Berdnik D, Shen J, Zou B, Xie X, Tingle M, Hinkson I, Angst M, Wyss-Coray T. Human umbilical cord plasma proteins revitalize hippocampal function in aged mice. Nature. [Internet] 2017; Apr 27;544(7651):488-492. [Cited 10 oct 2018]. Disponible en: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC558622240. Wang, L., Wang, D. Liu, M. Yao, R. Correlation between Tissue Characterization and Dynamic Expression of Matrix Metalloproteinase-2 and Its Tissue Inhibitor in Conjunctival Filtering Bleb of Rats. Biomed research international. [Internet] 2017; 2017, 10 pages. [Cited 16 Ene 2019]. Disponible en: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5624151/pdf/BMRI2017-1054129.pdf41. Gayatri, R., Crasta, J., Thomas, T., Pratibha, D., Thomas, A., Sridhar, T. S., Kurpad, A. V. Structural Analysis of the Umbilical Cord and Its Vessels in Intrauterine Growth Restriction and Pre-eclampsia. Journal of Fetal Medicine. [Internet]. 2017; 4(2), 85–92. [Cited 27 Abr 2019]. Disponible en: https://link.springer.com/article/10.1007/s40556-017-0118-242. Mottet, N., Chaussy, Y., Arbez-Gindre, F., & Riethmuller, D. Fisiología y patologías del cordón umbilical. EMC - Ginecología-Obstetricia. [Internet]. 2017; 53(4), 1–12. [Cited 17 May 2019]. Disponible en: https://www.sciencedirect.com/science/article/abs/pii/S1283081X1786887943. Sundrani D, Narang A, Mehendale S, Joshi S, Chavan-Gautam P. Investigating the expression of MMPs and TIMPs in preterm placenta and role of CpG methylation in regulating MMP-9 expression. IUBMB Journals. [Internet]. 2017; 69, 985-993 [Cited 18 Jul 2019]. Disponible en: https://iubmb.onlinelibrary.wiley.com/doi/full/10.1002/iub.168744. Zarkesh M, Zadeh-Vakili A, Azizi F, Fanaei SA, Foroughi F, Hedayati M. The Association of BRAF V600E Mutation With Tissue Inhibitor of Metalloproteinase-3 Expression and Clinicopathological Features in Papillary Thyroid Cancer. International Journal of Endocrinology and Metabolism. [Internet] 2018; 16(2): e56120. [Cited 16 oct 2018]. Disponible en: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5972213/45. Mashayekhi F, Saberi A, Mashayekhi S. Serum TIMP1 and TIMP2 concentration in patients with different grades of meningioma. Clinical Neurology and Neurosurgery. [Internet] 2018. 170, 84-87. [Cited 10 feb 2019]. Disponible en: https://www.sciencedirect.com/science/article/pii/S0303846718301756?via%3Dihub46. Liu Z, Ren Y, Zhu F. Expression of MMP-2 and TIMP‑3 with incidence and prognosis of giant-cell tumor of the bone. Oncology Letters. Oncol Lett. [Internet] 2018; 16 (1): 721–726. [Cited 16 oct 2018]. Disponible en: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6019976/47. R&D Systems. Quantikine ELISA - Human TIMP-2 Immunoassay. Catálogo DTM200 [Internet]. 2018. [Cited 18 mar 2019]. Disponible en: https://resources.rndsystems.com/pdfs/datasheets/dtm200.pdf48. Wang, W. Zhang, Y. Liu, M. Wang, Y. Yang, T. Li, D. Li, Q. TIMP2 is a Poor Prognostic Factor and Predicts Metastatic Biological Behavior in Gastric Cancer. Scientific Reports (Nature Publisher Group). [Internet]. 2018; 8, 1-11. [Cited 18 mar 2019]. Disponible en: https://ezproxy.unicolmayor.edu.co:2136/central/docview/2059010336/E4BDA59C6A134AE0PQ/1?accountid=5043849. R&D SYSTEMS a biotechne brand. Quantikine ELISA Human TIMP-2 Immunoassay [Internet]. USA R&D Systems, Inc; 2018. [Cited 16 mar 2019]. Disponible en: https://resources.rndsystems.com/pdfs/datasheets/dtm200.pdf50. Camacho, M., Escalona, R., Romero, S., Vargas, M., Zúñiga, M., Mora. J. Células madre de sangre de cordón umbilical: obtención, aplicaciones y situación en Costa Rica. REVISTA MÉDICA DE LA UNIVERSIDAD DE COSTA RICA. [Internet]. 2018; Vol 12 Nº 1. . [Cited 16 sep 2019]. Disponible en: https://revistas.ucr.ac.cr/index.php/medica/article/view/3461251. Schaduangrat, N, Prachayasittikul, V, Choomwattana, S, Wongchitrat, P, Phopin, K, Suwanjang, W, et al. Multidisciplinary approaches for targeting the secretase protein family as a therapeutic route for Alzheimer’s disease. Medicinal Research Reviews. [Internet] 2019. [Cited 16 Mar 2019]. Disponible en: https://www.ncbi.nlm.nih.gov/pubmed/3062809952. Medeiros, N. I., & Gomes, J. Cytometric Bead Array (CBA) for Measuring Cytokine Levels in Chagas Disease Patients. T. Cruzi Infection. [Internet] 2019; vol 1955 309–314. [Cited 29 Apr 2019]. Disponible en: https://link.springer.com/protocol/10.1007/978-1-4939-9148-8_2353. Malemud CJ. Inhibition of MMPs and ADAM/ADAMTS. Biochem Pharmacol. [Internet] 2019;165:33-40. [Cited 29 sep 2019]. Disponible en: https://www.ncbi.nlm.nih.gov/m/pubmed/30826330/54. Biolegend® Enabling Legendary Discovery™. Human TIMP Mix and Match Subpanel. [Internet] Vol 2. [Cited 29 sep 2018]. Disponible en: https://www.biolegend.com/Files/Images/media_assets/pro_detail/datasheets/75084_Hu_TIMP_Mix_and_Match_Subpanel_V2.pdfDerechos Reservados -Universidad Colegio Myor de Cundinamarca ,2019https://creativecommons.org/licenses/by-nc-sa/4.0/info:eu-repo/semantics/openAccessAtribución-NoComercial-CompartirIgual 4.0 Internacional (CC BY-NC-SA 4.0)http://purl.org/coar/access_right/c_abf2Cuantificación de TIMP-2 en plasmas de sangre de cordón umbilical de neonatos colombianos y su correlación con variables neonatales utilizando el método CBA en citometría de flujoTrabajo de grado - Pregradohttp://purl.org/coar/resource_type/c_7a1fhttp://purl.org/coar/version/c_970fb48d4fbd8a85Textinfo:eu-repo/semantics/bachelorThesishttps://purl.org/redcol/resource_type/TPinfo:eu-repo/semantics/publishedVersionBogotá D.C.Medicina regenerativaIngeniería de tejidosEmbriologíaTIMP-2CBAAlzheimerhipocampoPlasma de sangre de cordón umbilicalORIGINALSustentación - TIMP-2.pdfSustentación - TIMP-2.pdfapplication/pdf409622https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/214/1/Sustentaci%c3%b3n%20-%20TIMP-2.pdfd5149307b9ac41f611913f07cea31ae0MD51open access1 PARA SUBIR Tesis - TIMP-2.pdf1 PARA SUBIR Tesis - TIMP-2.pdfapplication/pdf1112773https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/214/2/1%20PARA%20SUBIR%20Tesis%20-%20TIMP-2.pdf157f1da9550ed17e43c700a4cc45ef38MD52open access2 PARA SUBIR Tesis - TIMP-2 (1).pdf2 PARA SUBIR Tesis - TIMP-2 (1).pdfapplication/pdf59024https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/214/3/2%20PARA%20SUBIR%20Tesis%20-%20TIMP-2%20%281%29.pdfca4206b2f79670f656fe0deb8d080854MD53metadata only accessLICENSElicense.txtlicense.txttext/plain; charset=utf-814828https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/214/4/license.txt2f9959eaf5b71fae44bbf9ec84150c7aMD54open accessTEXTSustentación - TIMP-2.pdf.txtSustentación - TIMP-2.pdf.txtExtracted texttext/plain16156https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/214/5/Sustentaci%c3%b3n%20-%20TIMP-2.pdf.txtc8753b61349839bd0633274e6fa698afMD55open access1 PARA SUBIR Tesis - TIMP-2.pdf.txt1 PARA SUBIR Tesis - TIMP-2.pdf.txtExtracted texttext/plain106960https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/214/7/1%20PARA%20SUBIR%20Tesis%20-%20TIMP-2.pdf.txtd00c31b5323af4579136df57a96e9af9MD57open access2 PARA SUBIR Tesis - TIMP-2 (1).pdf.txt2 PARA SUBIR Tesis - TIMP-2 (1).pdf.txtExtracted texttext/plain2https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/214/9/2%20PARA%20SUBIR%20Tesis%20-%20TIMP-2%20%281%29.pdf.txte1c06d85ae7b8b032bef47e42e4c08f9MD59metadata only accessTHUMBNAILSustentación - TIMP-2.pdf.jpgSustentación - TIMP-2.pdf.jpgGenerated Thumbnailimage/jpeg6947https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/214/6/Sustentaci%c3%b3n%20-%20TIMP-2.pdf.jpgc0ff83d31576e4ff81eb95c8eddc6d45MD56open access1 PARA SUBIR Tesis - TIMP-2.pdf.jpg1 PARA SUBIR Tesis - TIMP-2.pdf.jpgGenerated Thumbnailimage/jpeg6639https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/214/8/1%20PARA%20SUBIR%20Tesis%20-%20TIMP-2.pdf.jpg5fb0bd793d9aacad85f82745fe98a231MD58open access2 PARA SUBIR Tesis - TIMP-2 (1).pdf.jpg2 PARA SUBIR Tesis - TIMP-2 (1).pdf.jpgGenerated Thumbnailimage/jpeg6054https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/214/10/2%20PARA%20SUBIR%20Tesis%20-%20TIMP-2%20%281%29.pdf.jpg8f62c0a3866786294521b43bb5c58a0cMD510metadata only accessunicolmayor/214oai:repositorio.unicolmayor.edu.co:unicolmayor/2142021-06-11 03:00:14.735An error occurred on the license name.|||https://creativecommons.org/licenses/by-nc-sa/4.0/open accessBiblioteca Digital Unicolmayorrepositorio@unicolmayor.edu.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 |