CAJ | 학술논문

背景：如何获得大量稳定活力好的细胞是人脐带间充质干细胞培养的难点。 　　目的：通过组织块法、酶消化法和酶解组织块法3种细胞培养方法比较，筛选出最佳的人脐带间充质干细胞的培养方式。 　　方法：分离新鲜脐带10根，分别采用组织块法、酶消化法、酶解组织块法3种方式培养人脐带间充质干细胞，比较3种方式原代人脐带间充质干细胞爬出所需时间、细胞培养成功率、绘制细胞增殖曲线，利用流式细胞仪检测细胞表面标志，并进行多项分化能力检测。 　　结果与结论：酶解组织块培养法原代人脐带间充质干细胞爬出所需时间与酶消化法无显著差异，但明显短于组织块法(P <0.01)，原代人脐带间充质干细胞培养成功率显著高于其他两组，人脐带间充质干细胞增殖情况3组无显著性差异，酶解组织块培养法培养的第3代人脐带间充质干细胞其细胞表面标志及多项分化能力符合间充质干细胞的特性。酶解组织块培养法能缩短原代人脐带间充质干细胞爬出时间，显著提高原代人脐带间充质干细胞的成功率。
배경：여하획득대량은정활력호적세포시인제대간충질간세포배양적난점。 　　목적：통과조직괴법、매소화법화매해조직괴법3충세포배양방법비교，사선출최가적인제대간충질간세포적배양방식。 　　방법：분리신선제대10근，분별채용조직괴법、매소화법、매해조직괴법3충방식배양인제대간충질간세포，비교3충방식원대인제대간충질간세포파출소수시간、세포배양성공솔、회제세포증식곡선，이용류식세포의검측세포표면표지，병진행다항분화능력검측。 　　결과여결론：매해조직괴배양법원대인제대간충질간세포파출소수시간여매소화법무현저차이，단명현단우조직괴법(P <0.01)，원대인제대간충질간세포배양성공솔현저고우기타량조，인제대간충질간세포증식정황3조무현저성차이，매해조직괴배양법배양적제3대인제대간충질간세포기세포표면표지급다항분화능력부합간충질간세포적특성。매해조직괴배양법능축단원대인제대간충질간세포파출시간，현저제고원대인제대간충질간세포적성공솔。
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.