人妊娠子宫平滑肌细胞培养消化法与组织块法效果比较

The effect comparison of human pregnant uterine smooth muscle cells culture methods between digestion and tissue culture

目的寻求简单有效的人妊娠子宫平滑肌细胞原代培养方法,建立稳定、高纯度、高产量的子宫平滑肌细胞原代培养体外模型。方法利用20例妊娠子宫组织,分别使用组织块法和胶原酶消化法分离及纯化人子宫平滑肌细胞,通过免疫荧光方法检测平滑肌细胞的肌动蛋白(α-SMA)及钙调节蛋白(calponin)的表达。结果组织块法和胶原酶消化法培养的平滑肌细胞均呈梭型,峰谷样生长,α-SMA及calponin的免疫荧光化学检测结果为阳性,胶原酶消化法较组织块培养法稳定且缩短原代培养时间。结论胶原酶法简便,快速,稳定,可建立稳定、高纯度的子宫平滑肌细胞体外培养体系。

Objective To search for a simple and effective primary culture method for human uterine smooth muscle ceils, and establish a stable, high purity and high quantity of the uterine smooth muscle cells in primary culture model in vitro. Methods 20 cases of uterine tissue respectively used tissue explant and collagenase digestion method for isolation and purification of human uterine smooth muscle cells. The smooth muscle cells were detected by immunofluores-eenee method of α - SMA and calponin expression. Results Tissue method and collagenase digestion method in cultu- ring smooth muscle cells showed shuttle type, peak-valley-like growth. The immunefluorescenee chemical test results of α -SMA and calponin were positive. But the collagenase digestion method is more stable and shorter than the method of tissue culture of primary culture time. Conclusion The collagenases digestion method is simple, rapid and stable, and can establish a stable high purity of the uterine smooth muscle cell culture system in vitro.