Damjanov A, Wewer U M, Tuma B, Damjanov I
Department of Pathology and Cell Biology, Jefferson Medical College, Thomas Jefferson University, Philadelphia, PA 19107.
Differentiation. 1990 Nov;45(2):84-95. doi: 10.1111/j.1432-0436.1990.tb00461.x.
Three new cell lines (NE, ME, LRD) were cloned from mouse-embryo-derived teratocarcinomas and characterized on the basis of developmental, ultrastructural, and cytochemical criteria as nullipotent embryonal carcinoma (EC), pure parietal yolk sac (PYS) carcinoma and mixed parieto-visceral yolk sac carcinoma respectively. Cell lines NE and ME were composed of a monomorphous cell population; however, the morphology of ME was growth-medium-dependent. LRD was composed of a heterogeneous cell population and formed embryoid bodies. NE secreted soluble laminin, osteonectin, entactin and fibronectin but did not form visible pericellular matrix. ME formed pericellular matrix which was composed of laminin and entactin, but did not contain fibronectin. The LRD cells formed pericellular matrix which was composed of laminin, entactin and fibronectin. Whereas laminin from ME and LRD reacted with polyclonal antibodies and a monoclonal antibody to parietal yolk sac laminin, the laminin from NE cells was unreactive with the monoclonal antibody. Osteonectin was found in the supernatant of LRD and ME, but could not be demonstrated immunohistochemically in the extracellular matrix. We conclude that some extracellular matrix components, such as laminin and fibronectin, are produced not only by yolk sac carcinoma cells but by nullipotent EC as well, although the latter do not assemble them into extracellular matrix. Laminin produced by EC is immunochemically different from laminin secreted by yolk sac carcinoma. The extracellular matrix produced by mixed parieto-visceral yolk sac carcinoma is different from the matrix laid down by the pure PYS in that the latter does not contain fibronectin. The lack of osteonectin in the extracellular matrix of yolk sac carcinoma cells indicates that not all polypeptides secreted by these cell lines are incorporated into the extracellular matrix. The new cell lines described in this paper differ with regard to their capacity to form extracellular matrix and secrete its various components. Hence they could be used for further studies of basement membrane assembly in vitro.
从源自小鼠胚胎的畸胎瘤中克隆出三种新的细胞系(NE、ME、LRD),并根据发育、超微结构和细胞化学标准分别鉴定为无潜能胚胎癌(EC)、纯壁层卵黄囊(PYS)癌和混合壁 - 脏层卵黄囊癌。细胞系NE和ME由单一形态的细胞群体组成;然而,ME的形态依赖于生长培养基。LRD由异质性细胞群体组成并形成胚状体。NE分泌可溶性层粘连蛋白、骨连接蛋白、巢蛋白和纤连蛋白,但不形成可见的细胞周基质。ME形成由层粘连蛋白和巢蛋白组成但不含纤连蛋白的细胞周基质。LRD细胞形成由层粘连蛋白、巢蛋白和纤连蛋白组成的细胞周基质。虽然来自ME和LRD的层粘连蛋白与多克隆抗体和针对壁层卵黄囊层粘连蛋白的单克隆抗体发生反应,但来自NE细胞的层粘连蛋白与该单克隆抗体无反应。在LRD和ME的上清液中发现了骨连接蛋白,但在细胞外基质中无法通过免疫组织化学方法证实。我们得出结论,一些细胞外基质成分,如层粘连蛋白和纤连蛋白,不仅由卵黄囊癌细胞产生,也由无潜能的EC产生,尽管后者不会将它们组装成细胞外基质。EC产生的层粘连蛋白在免疫化学上与卵黄囊癌分泌的层粘连蛋白不同。混合壁 - 脏层卵黄囊癌产生的细胞外基质与纯PYS形成的基质不同,因为后者不含纤连蛋白。卵黄囊癌细胞的细胞外基质中缺乏骨连接蛋白表明并非这些细胞系分泌的所有多肽都被整合到细胞外基质中。本文描述的新细胞系在形成细胞外基质和分泌其各种成分的能力方面存在差异。因此,它们可用于体外进一步研究基底膜组装。