人胎儿毛囊隆突细胞的分离、培养与鉴定

Isolation, cultivation and identification of human fetal follicular bulge cells in vitro

目的：建立一种简单高效的毛囊隆突细胞的培养方法．方法：将5～7mo终止妊娠胎儿头皮的毛干中上部剪下后Ⅰ型胶原酶消化，采用成纤维细胞和毛囊隆突细胞对胰酶的不同敏感性进行纯化，流式细胞仪进行细胞周期分析，MTT法检测细胞克隆率，免疫组化检测细胞K19的表达情况．结果：消化法原代培养可以每小时获得100个左右毛囊隆突，经姨酶消化不同时间可以纯化毛囊隆突细胞，流式细胞仪检测第二代细胞Gl期占88．20％，细胞克隆形成率为18．2％，K19表达呈阳性．结论：建立了一种简单高效的毛囊隆突细胞的分离、培养方法，并证明其具有某些干细胞的特征性表现．

AIM： To develop a rapid and reproducible method for the culture of pure follicular bulge cells. METHODS ： 5-7 month fetal follicular bulge was isolated and incubated in collagenase I for 6 - 8 h, and bulge cells were purified using trypsin, identified using immunocytochemical methods;flow cytometry and MTT assay were applied to measure the clone rate and cell cycle. RESULTS ： About 100 follicular bulges were obtained each hour, and the bulge cells were purified by trypsin. The cell percentage in phase G1 was 88.20%, the clone formation rate was 18.2% and the K19 positive cells could be detected. CONCLUSION： A rapid and reproducible method has been developed for the culture of pure follicular bulge cells which represent many characters of some stem cells.