Xin yixue (Dec 2023)

Effects of different serum-free culture systems on the biological function of hUC-MSCs

  • Lin Huizhu, Li Cuiping, Chen Xiaoyan, Yang Li

DOI
https://doi.org/10.3969/j.issn.0253-9802.2023.12.005
Journal volume & issue
Vol. 54, no. 12
pp. 864 – 871

Abstract

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Objective To evaluate the effects of three different serum-free culture systems on the biological function of human umbilical cord mesenchymal stem cells （hUC-MSC） cultured in vitro. Methods Three strains of umbilical cord tissues were obtained，divided into 3 experimental groups and cultured to the 3rd generation （P3） in three different serum-free culture systems （Group A： serum-free culture system synthesized by pure chemical components，group B： serum-free culture system consisting of platelet lysate+basic culture medium，group C： serum-free culture system consisting of fibrinogen coating+recombinant human protein）. The cell proliferation，cell phenotype，three-line differentiation and immunomodulatory function were compared. Results All the cells in three groups were long and flat fusiform，adherent to the wall and uniform in size. The proliferation curve trend was similar，and the proliferation ability of P2 and P3 generations in groups A and B was better than that in group C （both P <0.05）. The cells in three experimental groups all met the requirements of the International Stem Cell Society for identification of hUC-MSC cell surface markers. The expression levels of CD29 and CD90 in P3 generations in groups A and B were higher than those in group C （all P < 0.05），and all three experimental groups had the ability of three-line differentiation. Compared with the control group，the proliferation rate of T lymphocytes in P3 generation and the proportion of cytokines secreted by Th1 and Th17 cell subsets were significantly decreased in the three experimental groups，and the proportion of regulatory T cells （CD4+ CD25+ FOXP3+） was significantly increased in the three experimental groups，but there was no statistical significance among three experimental groups （all P > 0.05）.Conclusions The hUC-MSC in three groups can maintain basic biological characteristics and immunomodulatory activity. The proliferation ability and expression levels of cell surface markers in groups A and B are superior to those in group C.

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