CAJ | 학술논문

目的：开发一种新的培养人胚胎干细胞（ hESCs）的包被基质，使hESCs的培养更加简便。方法用甲醇固定的小鼠胚胎成纤维细胞（ MEF）作为包被基质，人胚胎干细胞系X-01在该基质上生长，每隔5～6 d传代一次，培养10代后，对人胚胎干细胞特性进行检测，包括细胞形态、碱性磷酸酶染色、相关多能性基因的表达和分化能力。结果 hESCs在新的基质上生长良好，经10次传代后仍能保持典型的hESCs克隆形态。碱性磷酸酶染色阳性，免疫荧光染色 Oct4、SSEA4、Tra-1-60为阳性，体外分化可形成拟胚体。结论此种固定的基质可以大量制备，长期保存，并可以长期维持hESCs的未分化状态，为人胚胎干细胞的体外扩增探索出了一个新的途径。
목적：개발일충신적배양인배태간세포（ hESCs）적포피기질，사hESCs적배양경가간편。방법용갑순고정적소서배태성섬유세포（ MEF）작위포피기질，인배태간세포계X-01재해기질상생장，매격5～6 d전대일차，배양10대후，대인배태간세포특성진행검측，포괄세포형태、감성린산매염색、상관다능성기인적표체화분화능력。결과 hESCs재신적기질상생장량호，경10차전대후잉능보지전형적hESCs극륭형태。감성린산매염색양성，면역형광염색 Oct4、SSEA4、Tra-1-60위양성，체외분화가형성의배체。결론차충고정적기질가이대량제비，장기보존，병가이장기유지hESCs적미분화상태，위인배태간세포적체외확증탐색출료일개신적도경。
Objective To reduce the animal component contamination for human embryonic stem cells ( hESCs ) and to simplify hESCs culture process , we develop a new coating substrate which can support the hESCs growth without dif -ferentiation, and is easy to store and use. Methods Mouse embryonic fibroblasts(MEF)were fixed on the surface of plate by methanol.hESCs were cultured on this new substrate and were passaged every 5 to 6 days.After 10 passages, we checked the cell morphology , alkaline phosphatase expression , embryonic specific markers and the differentiation ability in vitro.Results After 10 passages , the hESCs grew well on this new substrate and maintained the typical hESCs morpholo -gy.Alkaline phosphatase staining was positive .Immunofluorescence staining showed that the expressions of Oct 4, SSEA4, Tra-1-60 were positive .The cells formed embryoid body in vitro .Conclusions This methanol-fixed MEF substrate can support the growth of undifferentiated hESCs .The coating material can be produced in large scale and stored for a long time.It provides a new and relatively easy way to amplify hESCs .