Extracción, cultivo y caracterización de células mesenquimales de médula ósea en biodispositivos para la regeneración del miocardio infartado

Currently tissue engineering strategies for myocardial regeneration after infarction are explored, including scaffolds that offer mechanical support and cell delivery into the injury. Bone marrow mesenchymal stem cells (MSC) are important candidates for cell therapy due to its ability to differentia...

Full description

Autores:
Melo Escobar, María Isabel
Tipo de recurso:
Trabajo de grado de pregrado
Fecha de publicación:
2019
Institución:
Universidad Autónoma de Occidente
Repositorio:
RED: Repositorio Educativo Digital UAO
Idioma:
spa
OAI Identifier:
oai:red.uao.edu.co:10614/10909
Acceso en línea:
http://hdl.handle.net/10614/10909
Palabra clave:
Ingeniería Biomédica
Ingeniería de tejidos
Scaffold
Infarto del miocardio
Células madre mesenquimales
Biomaterial
Viabilidad celular
Migración celular
Medicina regenerativa
Rights
openAccess
License
Derechos Reservados - Universidad Autónoma de Occidente
Description
Summary:Currently tissue engineering strategies for myocardial regeneration after infarction are explored, including scaffolds that offer mechanical support and cell delivery into the injury. Bone marrow mesenchymal stem cells (MSC) are important candidates for cell therapy due to its ability to differentiate into cells of cardiac tissue. However, the underlying mechanisms of MSC to promote tissue regeneration are not fully understood. The present study examined the undifferentiated and differentiated MSC’s behavior on a biopolymer, to assess cell viability and cell migration. The MSC were isolated from Wistar rats aged between 4 and 8 weeks. An improved isolation protocol was executed to optimize the performance of the cells in the scaffold. Group 1 (G1) of scaffolds (750 cells/µL) and group 2 (G2) (5000 cells/µL) were studied through trypan blue exclusion test to compare cell viability during 4 weeks. To assess cell migration group 3 (G3) were cell-seeded in a homogenous distribution and group 4 (G4) in a divided distribution, both at the same cell concentration of 2250 cells/µL. Cell migration was estimated through fluorescent microscopy. The isolation and cell culture protocol resulted in optimum confluence (>90%) in passage 4 to seed all the scaffolds. The cell viability assay determined G1 live cells had an average viability percentage of 98.23 ± 3.35 and for G2 an average of 98.38 ± 1.95. Distances measured in cell migration resulted highly similar (cv<1%). MSC showed optimal behavior during culture and differentiation and should be considered as good candidates for tissue regeneration. Their viability was significantly high, and it was not affected by the concentration of cells in the scaffold, the gelation method with ammonium hydroxide, the use of PETG in 3D printing or the integration to the biopolymer. Closeness in the distances evaluated between cellular reference points for cell migration, showed that there was no significant cellular migration. This suggests that cells did not generate sufficient tensile forces to create focal adhesions in the scaffold. Despite the favorable characteristics of MSC it is important to extend the study by modifying the biopolymer and submitting cellular constructs to paracrine factors of the natural myocardial infarcted microenvironment