Résumé
BACKGROUND:The studies have shown that the mesenchymal stem cel s derived from bone marrow and umbilical cord can be continuously cultured in vitro, and maintain the characteristics of stem cel s. The mesenchymal stem cel s can differentiate into hepatocyte-like cel s after“cocktail”induction by various cytokines. OBJECTIVE:To further identify whether umbilical cord-derived mesenchymal stem cel s in vitro co-cultured with normal hepatocytes can differentiate into hepatocyte-like cel s, and to investigate the differentiation method. METHODS:Mesenchymal stem cel s were isolated from human umbilical cord with adherent method, and the surface markers of umbilical cord-derived mesenchymal stem cel s were detected with flow cytometry. The umbilical cord-derived mesenchymal stem cel s were co-cultured with liver LO2 cel s 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 cel s could isolated from human umbilical cord successful y, showing fibroblastic morphology and adherent cel characterization. Among these cel s, 96.02%cel s were CD29 positive cel s and 96.6%cel s were CD105 positive cel s. The percentage of CD34 negative cel s was 99.65%. The percentage of CD105+CD29+double positive cel s was 94.84%. The mRNA of alpha-fetoprotein was found on the 7th day after co-cultured with LO2 cel s, 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 cel s can differentiate into hepatocyte-like cel s after co-cultured with normal hepatocytes.