Godkin J D, Bazer F W, Roberts R M
J Reprod Fertil. 1985 Jul;74(2):377-82. doi: 10.1530/jrf.0.0740377.
Primary cell cultures were established from cells derived from dissociated Day-14 and -16 sheep and pig blastocysts. The appearance of cells in culture from both species was similar. Cultures contained a variety of cells with distinct morphologies, some were small and compact and formed clumps and multiple layers while others were large, flat and formed a monolayer. Within 4 h of culturing small floating fluid-filled spheres of cells were observed in the medium; some of these increased in size to greater than 1 cm diameter over 1-2 weeks. In addition, fluid-filled domes of cells arose from the underlying monolayer. Contractile cells became evident after about 8 days and some became organized into large patches of contracting tissue. Two-dimensional polyacrylamide gel electrophoresis and fluorography were performed on proteins released into the medium by confluent monolayers, floating spheres and floating cells that failed to attach during the first 24 h. All cultures produced as major products proteins with electrophoretic mobilities identical to certain fetal plasma proteins. In general, cultures did not produce proteins characteristic of short-term cultures of whole conceptuses harvested at Days 14-16. In cultures established from sheep blastocysts only the cells that failed to attach produced ovine trophoblast protein-1, a major polypeptide produced by the trophectoderm of the sheep conceptus between Days 13 and 21 of pregnancy.
原代细胞培养是从第14天和第16天的绵羊和猪囊胚解离得到的细胞建立的。两种物种培养的细胞外观相似。培养物中含有多种形态各异的细胞，有些细胞小而紧密，形成团块和多层，而另一些细胞大而扁平，形成单层。培养4小时内，在培养基中观察到小的漂浮的充满液体的细胞球；其中一些在1 - 2周内直径增大到超过1厘米。此外，细胞的充满液体的穹顶从下面的单层细胞中出现。约8天后收缩细胞变得明显，一些收缩细胞组织成大片收缩组织。对汇合单层细胞、漂浮球以及在最初24小时内未能附着的漂浮细胞释放到培养基中的蛋白质进行了二维聚丙烯酰胺凝胶电泳和荧光自显影分析。所有培养物产生的主要产物是电泳迁移率与某些胎儿血浆蛋白相同的蛋白质。一般来说，培养物不产生第14 - 16天收获的完整概念体短期培养物特有的蛋白质。在从绵羊囊胚建立的培养物中，只有未能附着的细胞产生绵羊滋养层蛋白-1，这是妊娠第13至21天期间绵羊概念体滋养外胚层产生的一种主要多肽。
