Human umbilical cord-derived mesenchymal stem cells co-cultured with hepatocytes can differentiate into hepatocyte-like cells

doi:10.3969/j.issn.2095-4344.2013.32.005

Abstract

Abstract:

BACKGROUND: The studies have shown that the mesenchymal stem cells derived from bone marrow and umbilical cord can be continuously cultured in vitro, and maintain the characteristics of stem cells. The mesenchymal stem cells can differentiate into hepatocyte-like cells after “cocktail” induction by various cytokines.
OBJECTIVE: To further identify whether umbilical cord-derived mesenchymal stem cells in vitro co-cultured with normal hepatocytes can differentiate into hepatocyte-like cells, and to investigate the differentiation method.
METHODS: Mesenchymal stem cells were isolated from human umbilical cord with adherent method, and the surface markers of umbilical cord-derived mesenchymal stem cells were detected with flow cytometry. The umbilical cord-derived mesenchymal stem cells were co-cultured with liver LO2 cells without adding exogenous inducers. The expressions of alpha-fetoprotein, albumin and human cytokeratin 19 mRNA of hepatocyte specific markers were detected with reverse transcription PCR at 7, 14 and 21 days after culture, and periodic acid-Schiff staining was used to identify the functions.
RESULTS AND CONCLUSION: Mesenchymal stem cells could isolated from human umbilical cord successfully, showing fibroblastic morphology and adherent cell characterization. Among these cells, 96.02% cells were CD29 positive cells and 96.6% cells were CD105 positive cells. The percentage of CD34 negative cells was 99.65%. The percentage of CD105⁺CD29⁺ double positive cells was 94.84%. The mRNA of alpha-fetoprotein was found on the 7th day after co-cultured with LO2 cells, and the mRNA of albumin and human cytokeratin 19 were found on the 14th day. After co-cultured for 21 days, the alpha-fetoprotein mRNA could not be observed in the co-culture group. The expressions of albumin and human cytokeratin 19 were increased at 14 days. After co-cultured for 21 days, the glycogen staining was positive. Umbilical cord-derived mesenchymal stem cells can differentiate into hepatocyte-like cells after co-cultured with normal hepatocytes.

Key words: stem cells, umbilical cord/umbilical blood stem cells, umbilical cord-derived mesenchymal stem cells, hepatocytes, co-culture, cell differentiation, cell culture, hepatocyte-like cells, alpha-fetoprotein, albumin, human cytokeratin 19, provincial grants-supported paper, stem cell photographs-containing paper

CLC Number:

R394.2

Li Hua, Wen Feng, Qi Zhong-chun, Zhou Jin-jun, Zhu Ya-jie, Cheng Peng, Wei Dong, Su Xiao-mei, Tan Yong, Peng Jing-jing, Luo Qiao-li, Li Dong, Zhang Tao. Human umbilical cord-derived mesenchymal stem cells co-cultured with hepatocytes can differentiate into hepatocyte-like cells[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(32): 5772-5777.

Figures/Tables 4

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