人汗腺细胞的分离和纯化培养方法探讨

Isolation and purification of eccrine sweat glands in human skin

目的 探讨建立人皮肤汗腺细胞体外分离及纯化培养的技术。方法 通过酶消化法分离人皮肤汗腺 ,微量加样器在倒置显微镜屏视下吸取游离的汗腺组织移入 Hank平衡盐溶液 (HBSS) ,经纯化后予以原代培养。结果 分离的汗腺可在体外贴壁生长、增殖并传代 ,纯化处理清除了杂质细胞和组织碎片 ,解决了成纤维细胞对汗腺细胞培养的污染难题 ,且汗腺的生长能力无明显改变。结论 人汗腺细胞的培养存在着分离困难和成纤维细胞污染等难题 ,采用本方法可以获得大量处于良好生长状态的高纯度人汗腺细胞。

Objective To investigate the method of isolation and purification of epithelial cells of human eccrine sweat gland in vitro. Methods Through digesting human skin with collagenase type Ⅱ, cells of human eccrine sweat gland were isolated. Highly purified gland cells were obtained through transferring into the conditioned medium with a micropipette for at least three times under an inverted microscope. Primary culture was started immediately after purification. Cells could be further purified by enzyme digestion to eradicate the fibroblasts. Results Collagenase type Ⅱ could digest dermal collagen with little damage to gland cells. Isolated cells from human sweat glands were adherent to the wall of culture flask, and they grew well in cultures. The problem of contamination by tissue debris and other cells such as fibroblast could be overcome. Conclusion Isolation and purification of human sweat gland cells in vitro are still facing tough problems. Gland cells are successful to isolate and cultivate without contamination.