Culture methods for primary human umbilical cord mesenchymal stem cells / 中国组织工程研究

ABSTRACT

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