两种方法培养人早孕绒毛滋养层细胞的比较

Comparison of two ways to culture human primary first-trimester trophoblast cells

背景:人早孕绒毛滋养层细胞体外培养是研究各种妊娠疾病的基础,如何获得纯度高、数量多的滋养层细胞以及如何简化实验步骤,仍是目前研究的热点。目的:寻求一种培养高纯度人早孕绒毛滋养层细胞方法。方法:取5～10周正常绒毛组织,胰酶消化和差速贴壁联合应用法与组织块培养法分别进行人早孕绒毛滋养层细胞原代培养,免疫组织化学观察细胞角蛋白7和波形蛋白的表达,进行滋养层细胞纯度的鉴定。结果与结论:两种方法均能培养出人早孕绒毛滋养层细胞,呈三角形,多边形。抗-细胞角蛋白7阳性表达,抗-波形蛋白阴性表达。胰酶消化和差速贴壁联合应用法培养细胞纯度高于组织块培养法(P<0.05)。提示胰酶消化和差速贴壁联合应用法是一种培养人早孕绒毛滋养层细胞较好的方法。

BACKGROUND：In vitro culture of human primary first-trimester trophoblast cells is the study basis for various diseases in pregnancy.Currently,it is a hotspot to obtain a large amount of high-purity trophoblast cells as well as to simplify the experimental procedure.OBJECTIVE：To explore a culture method to harvest high-purity human primary first-trimester trophoblast cells.METHODS：Normal chorionic villi（5-10 weeks） were obtained for primary culture of human primary first-trimester trophoblast cells using tissue-culture method and trypsin digestion combined with differential adhesion method.We observed the expression of cytokeratin and vimentin by immunohistochemistry to identify the purity of cultured trophoblast cells.RESULTS AND CONCLUSION：These two methods could obtain human primary first-trimester trophoblast cells in triangular and polygonal shape.The positive expression of cytokeratin and negative expression of vimentin were found by immunohistochemistry.The purity of cultured cells using trypsin digestion combined with differential adhesion method was higher than that using tissue-culture method（P 0.05）.Trypsin digestion combined with differential adhesion method is a better method to culture human primary first-trimester trophoblast cells.