Department of Chemical Engineering, Faculty of Engineering, Kyushu University, 744 Motooka, Nishi-ku, Fukuoka 819-0395, Japan.
J Biosci Bioeng. 2010 Nov;110(5):582-7. doi: 10.1016/j.jbiosc.2010.06.002. Epub 2010 Jul 2.
Conventionally, embryonic stem (ES) cells are cultured on a cell layer of mouse embryonic fibroblasts (MEFs) as feeder cells to support undifferentiated growth of ES cells. In this study, cell-cell interactions between mouse ES and feeder cells were artificially engineered via an epithelial cell adhesion molecule, E-cadherin, whose expression is considerable in ES cells. Mouse mesenchymal STO and NIH3T3 cells that were genetically engineered to express E-cadherin were used in ES cell cultures as feeder cells. ES cells cultured on the E-cadherin-expressing feeder cells maintained the expression of stem cell markers, alkaline phosphatase (AP), Oct3/4, Nanog and Sox2, and the efficiency of AP-positive colony formation was comparable to MEFs, and much better than parental STO and NIH3T3 cells. Furthermore, ES cells maintained on the E-cadherin-expressing feeder cells possessed the ability to differentiate into the three germ layers both in vitro and in vivo. The results indicated that E-cadherin expression in feeder cells could improve the performance of feeder cells, which may be further applicable to create new artificial feeder cell lines.
传统上，胚胎干细胞（ES 细胞）在饲养层细胞（MEFs）的细胞层上培养，以支持 ES 细胞的未分化生长。在这项研究中，通过上皮细胞黏附分子 E-钙黏蛋白对 ES 细胞和饲养层细胞之间的细胞-细胞相互作用进行了人工设计，E-钙黏蛋白在 ES 细胞中的表达相当丰富。用基因工程表达 E-钙黏蛋白的小鼠间充质 STO 和 NIH3T3 细胞被用作饲养层细胞进行 ES 细胞培养。在表达 E-钙黏蛋白的饲养层细胞上培养的 ES 细胞维持着干细胞标志物碱性磷酸酶（AP）、Oct3/4、Nanog 和 Sox2 的表达，AP 阳性集落形成的效率与 MEFs 相当，比亲本 STO 和 NIH3T3 细胞好得多。此外，在表达 E-钙黏蛋白的饲养层细胞上维持的 ES 细胞具有在体外和体内分化为三个胚层的能力。结果表明，饲养层细胞中 E-钙黏蛋白的表达可以改善饲养层细胞的性能，这可能进一步适用于创建新的人工饲养层细胞系。
