Cherny R A, Findlay J K
Medical Research Centre, Monash Medical Centre, Melbourne, Victoria, Australia.
Biol Reprod. 1990 Aug;43(2):241-50. doi: 10.1095/biolreprod43.2.241.
Epithelial and stromal cells from endometria of ovariectomized estradiol-treated Corriedale ewes were separated and purified after collagenase digestion. The separation method utilized differences in the speed and ease of detachment of cultured epithelial and stromal cells attached to plastic in response to brief trypsin exposure. Cells were characterized according to morphological, growth, and histochemical criteria. Contamination of each cell type with the other was less than 1%. Separated cells were grown on plastic or on Matrigel-coated Millicell inserts with nitrocellulose membranes. Transmission and scanning electron microscope analyses demonstrated the existence of tight junctions and prominent microvilli in the epithelial cultures on inserts but not on plastic. Asymmetrical secretion of prostaglandin F2 alpha (PGF2 alpha) and prostaglandin E (PGE) by epithelial cells provided further evidence of polarization. Epithelial cell secretion of PGF2 alpha was greater than that by stromal cells whereas PGE secretion by stromal cells was greater than that by epithelial cells. Epithelial secretion in the basal direction was approximately 4 and 3 times that of apical secretion for PGF2 alpha and PGE, respectively. The separation protocol provides pure populations of ovine endometrial epithelial and stromal cells and the cultured epithelial cells exhibit characteristics of in vivo morphology and polarized function.
对经雌二醇处理的去卵巢考力代母羊子宫内膜的上皮细胞和基质细胞进行胶原酶消化后进行分离和纯化。分离方法利用了培养的上皮细胞和基质细胞在短暂暴露于胰蛋白酶后附着于塑料上的脱落速度和难易程度的差异。根据形态学、生长和组织化学标准对细胞进行表征。每种细胞类型被另一种细胞类型污染的程度小于1%。分离的细胞在塑料或涂有基质胶的带有硝酸纤维素膜的Millicell小室上生长。透射电子显微镜和扫描电子显微镜分析表明，在带有小室的上皮培养物中存在紧密连接和明显的微绒毛，而在塑料上则不存在。上皮细胞对前列腺素F2α（PGF2α）和前列腺素E（PGE）的不对称分泌提供了极化的进一步证据。上皮细胞分泌的PGF2α大于基质细胞，而基质细胞分泌的PGE大于上皮细胞。对于PGF2α和PGE，上皮细胞向基底方向的分泌分别约为向顶端分泌的4倍和3倍。该分离方案提供了纯的绵羊子宫内膜上皮细胞和基质细胞群体，并且培养的上皮细胞表现出体内形态和极化功能的特征。
