A Method for Primary Culture of Human Eutopic Endometrial Stromal Cells

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(χ2=3.68,P=0.465).There was significant difference in digestion time,unilayer aggregation time and culture achievement rate between 4 groups(P0.05).The achievement rate of trypsinase cell culture was significantly lower than those of 3 types of collagenase(P0.05).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(P0.05),but shorter than that of type Ⅰ and Ⅳ(P0.05).Screen cloth separation was not significantly different from centrifugal in cell culture achievement rate and cell purity(P0.05),the cell unilayer aggregation time of the former(6.0±0.3)was shorter than that of the latter(6.5±0.4)(P0.05).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.