Cooper S, Pera M F
Institute of Cancer Research, Royal Marsden Hospital, Sutton, Surrey, UK.
Development. 1988 Dec;104(4):565-74. doi: 10.1242/dev.104.4.565.
Normal mesenchymal cells, normal epithelial cells and many transformed epithelial cells require serum attachment factors and extracellular matrix proteins for growth and differentiation in vitro, and recent evidence strongly supports a role for extracellular matrix molecules in the regulation of cell movement in vivo during early embryogenesis. We previously described the isolation and characterization of cell lines representative of three types of stem cells most commonly found in human adult testicular teratomas, namely embryonal carcinoma cells, yolk sac carcinoma cells resembling visceral endoderm and yolk sac carcinoma cells resembling parietal endoderm (endodermal sinus tumour cells). Of these three cell types, only endodermal sinus tumour cells, which show particularly malignant behaviour in vivo, have no serum requirement for attachment and growth in vitro. Supernatants from endodermal sinus tumour cells support the attachment of embryonal carcinoma cells in serum-free medium. We demonstrate here that endodermal sinus tumour cells, but not other cell types isolated from testicular teratomas, secrete the serum attachment protein, vitronectin (also known as serum-spreading factor, S-protein or epibolin), as well as fibronectin, laminin and type IV collagen, into serum-free medium. Purified vitronectin from medium conditioned by endodermal sinus tumour cells supported both attachment and spreading of embryonal carcinoma cells in vitro, whereas cells attached but did not spread properly on surfaces coated with fibronectin or laminin. Peptides containing the RGD cell recognition sequence common to many attachment proteins blocked attachment of endodermal sinus tumour cells to untreated tissue-culture plastic in serum-free medium. The results suggest a possible role for vitronectin in regulating cell motility and growth in early development, and in the invasion and spread of teratomas in vivo.
正常间充质细胞、正常上皮细胞以及许多转化的上皮细胞在体外生长和分化需要血清附着因子和细胞外基质蛋白，并且最近的证据有力地支持了细胞外基质分子在早期胚胎发育过程中体内细胞运动调节中的作用。我们之前描述了从人类成人睾丸畸胎瘤中最常见的三种干细胞类型所代表的细胞系的分离和特性，即胚胎癌细胞、类似于脏内胚层的卵黄囊癌细胞和类似于壁内胚层的卵黄囊癌细胞（内胚窦瘤细胞）。在这三种细胞类型中，只有在体内表现出特别恶性行为的内胚窦瘤细胞在体外附着和生长不需要血清。内胚窦瘤细胞的上清液在无血清培养基中支持胚胎癌细胞的附着。我们在此证明，内胚窦瘤细胞而非从睾丸畸胎瘤中分离出的其他细胞类型，会将血清附着蛋白玻连蛋白（也称为血清扩散因子、S蛋白或表皮调节素）以及纤连蛋白、层粘连蛋白和IV型胶原分泌到无血清培养基中。从内胚窦瘤细胞条件培养基中纯化的玻连蛋白在体外支持胚胎癌细胞的附着和铺展，而细胞在涂有纤连蛋白或层粘连蛋白的表面能附着但不能正常铺展。含有许多附着蛋白共有的RGD细胞识别序列的肽在无血清培养基中阻断了内胚窦瘤细胞与未处理的组织培养塑料的附着。这些结果表明玻连蛋白在早期发育中调节细胞运动性和生长以及在体内畸胎瘤的侵袭和扩散中可能发挥作用。
