RESUMEN
Cell senescence leads to a number of changes in the properties of mesenchymal stromal cells (MSCs). In particular, the number of damaged structures is increased producing negative effect on intracellular processes. Elimination of the damaged molecules and organelles occurs via autophagy that can be important in the context of aging. Cultivation under low oxygen level can be used as an approach for enhancement of MSC therapeutic properties and "slowing down" cell senescence. The goal of this work was to study some morphological and functional characteristics and expression of autophagy-associated genes during replicative senescence of MSCs under different oxygen concentration. The study revealed changes in the regulation of autophagy at the transcriptional level. Upregulation of the expression of autophagosome membrane growth genes ATG9A and ULK1, of the autophagosome maturation genes CTSD, CLN3, GAA, and GABARAPL1, of the autophagy regulation genes TP53, TGFB1, BCL2L1, FADD, and HTT was shown. These changes were accompanied by downregulation of IGF1 and TGM2 expression. Increase of the lysosomal compartment volume was observed in the senescent MSCs that also indicated increase of their degradation activity. The number of lysosomes was decreased following prolonged cultivation under low oxygen concentration (5%). The replicative senescence of MSCs under conditions of different oxygen levels led to the similar modifications in the expression of the autophagy-associated genes.
Asunto(s)
Autofagia , Hipoxia de la Célula , Senescencia Celular , Células Madre Mesenquimatosas , Células Cultivadas , Humanos , Lisosomas/metabolismo , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/metabolismoRESUMEN
Proliferative activity of mesenchymal stromal cells isolated from five sources (chorionic villi, Wharton's jelly, amnion, endometrium, and adipose tissue) was compared by flow cytometry and real-time PCR (by the content of mRNA of genes encoding of cell cycle regulators). Mesenchymal stromal cells derived from the endometrium demonstrated maximum stability and high proliferative potential.
Asunto(s)
Tejido Adiposo/citología , Amnios/citología , Vellosidades Coriónicas/crecimiento & desarrollo , Endometrio/citología , Células Madre Mesenquimatosas/citología , Gelatina de Wharton/citología , Adulto , Diferenciación Celular , Proliferación Celular , Células Cultivadas , Femenino , Citometría de Flujo , Humanos , Reacción en Cadena en Tiempo Real de la Polimerasa , Adulto JovenRESUMEN
Cell cultures isolated from human endometrium by enzyme digestion consisted of highly viable fibroblast-like mesenchymal cells expressing CD90, CD73, and CD105. During passage 1, the cultures contained a small fraction of cytokeratin-7(+) epithelial cells that disappeared by passage 2. The cultures from the endometrium could be induced to adipogenic, osteogenic and chondrogenic differentiation in vitro. These findings suggest that human endometrium is a convenient source of biomaterial for minimally invasive isolation of cultures that exhibit typical morphology and immunophenotypic profile of resident multipotent mesenchymal stromal cells retain high viability in vitro.
