Carnegie J A, Cabaca O
Department of Obstetrics & Gynecology, Loeb Institute of Medical Research, Ottawa Civic Hospital, Ottawa, Ontario, Canada.
Biol Reprod. 1991 Oct;45(4):572-80. doi: 10.1095/biolreprod45.4.572.
Reichert's membrane is a basement membrane deposited on the inner surfaces of rat and mouse trophectodermal (TE) cells beginning at the blastocyst stage of embryonic development that may play a role in the migration of the parietal endodermal (PE) cells to form an inner lining to the TE. The abilities of various glycoproteins present in Reichert's membrane to support PE cell migration and replication in vitro were examined by isolating inner cell masses (ICMs) from Day 5 rat blastocysts (Day 1 = day of vaginal plug) and culturing them (24-72 h) either on surfaces that had been precoated with collagen IV, fibronectin, or laminin or on thin (1-2-mm) gels of Matrigel (a tumor cell-derived basement membrane preparation) or type I collagen. Time-dependent changes in the area occupied by each ICM on the culture surface and the number of migrating cells per ICM were quantified by morphometric analysis. Type IV collagen, the basement membrane-specific collagen, supported ICM attachment and the outward migration (overall increase of approx. 60-fold in mean ICM area occupied on the culture surface) and proliferation (cell doublings following every 24 h of culture) of laminin-containing PE-like cells. These effects were not altered by the inclusion of exogenous fibronectin or laminin in the culture medium. Collagen IV coating concentrations as low as 0.16 micrograms/ml supported PE cell attachment and migration, and maximal responses were seen with a coating concentration of 0.63 micrograms/ml.(ABSTRACT TRUNCATED AT 250 WORDS)
赖歇特氏膜是一种基底膜,从胚胎发育的囊胚期开始沉积在大鼠和小鼠滋养外胚层(TE)细胞的内表面,可能在壁内胚层(PE)细胞迁移以形成TE的内衬中发挥作用。通过从第5天的大鼠囊胚(第1天=发现阴道栓的日子)中分离内细胞团(ICM),并将它们培养(24 - 72小时)在预先用IV型胶原、纤连蛋白或层粘连蛋白包被的表面上,或者在基质胶(一种肿瘤细胞衍生的基底膜制剂)或I型胶原的薄(1 - 2毫米)凝胶上,研究了赖歇特氏膜中存在的各种糖蛋白在体外支持PE细胞迁移和复制的能力。通过形态计量分析对每个ICM在培养表面占据的面积以及每个ICM迁移细胞的数量随时间的变化进行了量化。基底膜特异性胶原IV型支持ICM附着以及含层粘连蛋白的类PE细胞的向外迁移(培养表面上ICM平均占据面积总体增加约60倍)和增殖(培养每24小时细胞加倍)。培养基中加入外源性纤连蛋白或层粘连蛋白不会改变这些作用。低至0.16微克/毫升的IV型胶原包被浓度支持PE细胞附着和迁移,在包被浓度为0.63微克/毫升时观察到最大反应。(摘要截短于250字)