Abstract:

Objective To obtain mesenchymal stem cells(MSCs) from human umbilical cord blood (HUCB) in vitro and to study the condition in which MCSs differentiated into hepatocyte-like cells. Methods HUCB was collected after the baby was delivered, and mononuclear cells (MNCs) were separated by gradient centrifugation, and then using adherent culture, MSCs were obtained. CD29, CD44, CD34, CD45 expression levels were determined by flow cytometry.  Also the albumin (ALB), alpha-fetoprotein (AFP), cytokeration (CK19) mRNA expression levels were determined by RT-PCR. ResultsThe results of flow cytometry showed that CD29 (+) and CD44 (+) expressions were positive, however CD34 (-) and CD45 (-) expressions were negative. The adherent cells were proved to be MSCs. The group using several cytokines step by step showed the changes of cell morphology and expressions of AFPmRNA(+), ALBmRNA(+), and CK19mRNA(+) at week 8. But, the cell density was significantly decreased. Conclusion MSCs in HUCB are high-concentration-and highFCS(fetal culf serum) dependents.  MSCs could be differentiated into hepatocytes by an in vitro culture with several different cytokines step by step.

Key words: Human umbilical cord blood; Mesenchymal stem cells; Hepatocyte