中文摘要：

目的探讨子宫内膜异位症在位内膜间质细胞原代培养的最优方法。方法40例子宫内膜异位症患者（增殖期和分泌期内膜各20例）的在位内膜组织随机分为4组，分别采用胰蛋白酶、Ⅰ型胶原酶、Ⅱ型胶原酶、Ⅳ型胶原酶消化，随机选择离心分离或筛网分离间质细胞，行细胞免疫组织化学染色（XP法）鉴定。结果增殖期与分泌期内膜间质细胞培养成功率差异无统计学意义（Х^2=3．68，P〉0．05）。胰蛋白酶组的细胞培养成功率与其他3种胶原酶组的细胞培养成功率间差异有统计学意义（P〈0．05），3种胶原酶细胞培养成功率间差异无统计学意义（P〉0．05）；Ⅰ型胶原酶酶解所得间质细胞的单层汇聚时间[（5．0±0．8）d]最短（P〈0．05）；Ⅰ型胶原酶和胰蛋白酶的细胞消化时间差异无统计学意义（P〉0．05），但短于Ⅱ型和Ⅳ型胶原酶（P〈0．05）；筛网分离法与离心分离法在细胞培养成功率和细胞纯度上差异无统计学意义（P〉0．05），但前者的细胞单层汇聚时间[（6．0±0．3）d]短于后者[（6．5±0．4）d]（P〈0．05）。结论月经周期不影响细胞培养成功率；采用Ⅰ型胶原酶消化子宫内膜、筛网分离间质细胞是子宫内膜异位症在位内膜间质细胞培养的最好方法。

英文摘要：

Objective To approach the best method for primary culture of human eutopic endometrial stromal cells. Methods Eutopic endometrial tissues of 40 endometriosis patients （20 in proliferative phase, 20 in secretory ） were divided into 4 groups. The tissues were digested using trypsinase, collagenase types Ⅰ , Ⅱ, Ⅳ, respectively, and stromal cells were obtained randomly by centrifugal or screen cloth separation, and identified by immunohistochemical staining （SP method）. Results No significant difference was noted in achievement ratio of endometrial stromal cell culture between in proliferative phase and in secretory （ Х^2 = 3.68, P = 0. 465 ）. There was significant difference in digestion time, unilayer aggregation time and culture achievement rate between 4 groups （ P 〈 0. 05 ）. The achievement rate of trypsinase cell culture was significantly lower than those of 3 types of collagenase （ P 〈 0. 05 ）. Unilayer aggregation time of stromal cells obtained by collagenase type Ⅰ was the shortest （5.0 ±0. 8 d）. Collagenase Ⅰ was not significantly different from trypsinase in cell digestion time （P 〉0. 05）, but shorter than that of type Ⅰ and Ⅳ （ P 〈 0. 05 ）. Screen cloth separation was not significantly different from centrifugal in cell culture achievement rate and cell purity （P 〉 0. 05 ）, the cell unilayer aggregation time of the former （6. 0 ±0. 3 ） was shorter than that of the latter （6. 5 ±0. 4） （ P 〈 0. 05 ）. Conclusion Menstrual cycle has not influence on the achievement ratio of stromal cells. Using collagenase Ⅰ to digest endometrium and screen cloth to separate stromal cells is the best method for primary culture of eutopic endometrial stromal cells.