人卵巢癌干细胞球的分离与悬浮培养方法

目的：寻找分离培养卵巢癌干细胞球的简便方法。方法：用组织块培养及胶原酶消化法分离培养原代人卵巢癌细胞,分别采用高浓度血清和无血清悬浮培养3~5d后用Percoll液分离悬浮生长的肿瘤细胞球,机械性吹打细胞并进行传代悬浮培养约1个月,采用倒置显微镜观察并用肿瘤干细胞相关抗原（CD44,CD117和CD34）进行免疫细胞化学染色来寻找卵巢癌干细胞标记。结果：采用高浓度血清培养和无血清干细胞培养液均能得到卵巢癌干细胞球。采用胶原酶消化法得到的细胞球相对较多,CD44可能是卵巢癌干细胞较特异的抗原标记。结论：高浓度血清培养能获得一定数量的肿瘤干细胞球,用Percoll液分离纯化肿瘤细胞球是一种简便的好方法。卵巢癌干细胞球多表达CD44抗原标记。

Isolation and culture of ovarian cancer-initiating spheroid cells from primary human tumors

OBJECTIVE:To improve the culture methods of ovarian cancer-initiating spheroid cells. METHODS: The tumor spheroid cells were isolated by tissue culture and collagenase I digestion, then high concentration of serum and serum-free medium were used. Percoll solution was used to isolate tumor spheroid cells after 3 or 5 days. and it was then cultured in suspension about one month. Tumor spheroid cells were identified by light microscope and immunocytochemical method stained for CD44, CD117 and CD34. RESULTS: Bo...