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Title: Mesenchymal stem cells from umbilical cord blood: parameters for isolation, characterization and adipogenic differentiation
Authors: Sibov, Tatiana Tais
Severino, P.
Marti, L. C.
Pavon, L. F.
Oliveira, D. M.
Tobo, P. R.
Campos, A. H.
Paes, A. T.
Amaro, E.
Gamarra, L. F.
Moreira-Filho, C. A.
Hosp Israelita Albert Einstein
Universidade de Brasília (UnB)
Universidade Federal de São Paulo (UNIFESP)
Keywords: Mesenchymal stem cells
Umbilical cord blood
Cell isolation
Cell characterization and adipogenesis
Issue Date: 1-Oct-2012
Publisher: Springer
Citation: Cytotechnology. Dordrecht: Springer, v. 64, n. 5, p. 511-521, 2012.
Abstract: Isolation of mesenchymal stem cells (MSCs) from umbilical cord blood (UCB) from full-term deliveries is a laborious, time-consuming process that results in a low yield of cells. in this study we identified parameters that can be helpful for a successful isolation of UCB-MSCs. According to our findings, chances for a well succeeded isolation of these cells are higher when MSCs were isolated from UCB collected from normal full-term pregnancies that did not last over 37 weeks. Besides the duration of pregnancy, blood volume and storage period of the UCB should also be considered for a successful isolation of these cells. Here, we found that the ideal blood volume collected should be above 80 mL and the period of storage should not exceed 6 h. We characterized UCB-MSCs by morphologic, immunophenotypic, protein/gene expression and by adipogenic differentiation potential. Isolated UCB-MSCs showed fibroblast-like morphology and the capacity of differentiating into adipocyte-like cells. Looking for markers of the undifferentiated status of UCB-MSCs, we analyzed the UCB-MSCs' protein expression profile along different time periods of the differentiation process into adipocyte-like cells. Our results showed that there is a decrease in the expression of the markers CD73, CD90, and CD105 that correlates to the degree of differentiation of UCB-MSCs We suggest that CD90 can be used as a mark to follow the differentiation commitment degree of MSCs. Microarray results showed an up-regulation of genes related to the adipogenesis process and to redox metabolism in the adipocyte-like differentiated MSCs. Our study provides information on a group of parameters that may help with successful isolation and consequently with characterization of the differentiated/undifferentiated status of UCB-MSCs, which will be useful to monitor the differentiation commitment of UCB-MSC and further facilitate the application of those cells in stem-cell therapy.
ISSN: 0920-9069
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