Culture methods for primary human umbilical cord mesenchymal stem cells

BACKGROUND: How to get a lot of stable and dynamic human umbilical cord mesenchymal stem cells is still a difficulty. OBJECTIVE: To investigate the best culture way of human umbilical cord mesenchymal stem cells by comparing tissue explant, enzyme digestion, enzymolysis methods. METHODS: Ten fresh umbilical cords were isolated to human umbilical cord mesenchymal stem cells by using tissue explant, enzyme digestion, and enzymolysis methods, respectively. Hereafter, we compared cells harvested using three culture methods in terms of the time for the primary cells to creep, successful rates of cell-culturing, and curves of cell growth. Surface markers of cells were detected by flow cytometry and multiple differentiations of cells were detected. RESULTS AND CONCLUSION: The time for the primary cells to creep out in the enzymolysis and enzyme digestion groups had no significant difference, but both of them were significantly shorter than that in the tissue explant group (P < 0.01). The successful rate of primary cell culture in the enzymolysis group was significantly higher than that in the other two groups. The cell proliferations of three groups had no significant difference. The cell surface markers and differentiation ability of the third generation of human umbilical cord mesenchymal stem cells cultured by enzymolysis method met the characteristics of mesenchymal stem cells. In conclusion, the enzymolysis method can shorten the time for primary human umbilical cord mesenchymal stem cells to creep out and increase the successful rates of primary human umbilical cord mesenchymal stem cells.