CAJ | 학술논문

目的：探索人胚胎干细胞建系方法，为干细胞和再生医学领域的科学研究提供可靠的实验方法和技术。方法分别利用有饲养层和无饲养层培养体系培养人的胚胎干细胞，用免疫荧光法、逆转录聚合酶链式扩增（ RT -PCR）法和体内形成畸胎瘤的方法鉴定其干细胞特征。结果培养的人胚胎干细胞，传代至33代时表达干细胞表面标志物 Oct -4，SSEA -3，Tra -1-60，Tra -1-81和全能型因子 C - MYC KLF -4 OCT -4 SOX -2，畸胎瘤组织切片中分别有消化腺内皮（内胚层）、脂肪组织（中胚层）和神经组织（外胚层）等组织形成，说明该胚胎干细胞形成的畸胎瘤有向三胚层分化的潜能。结论成功建立了人胚胎干细胞建系，所建立的培养人胚胎干细胞的方法可行。
목적：탐색인배태간세포건계방법，위간세포화재생의학영역적과학연구제공가고적실험방법화기술。방법분별이용유사양층화무사양층배양체계배양인적배태간세포，용면역형광법、역전록취합매련식확증（ RT -PCR）법화체내형성기태류적방법감정기간세포특정。결과배양적인배태간세포，전대지33대시표체간세포표면표지물 Oct -4，SSEA -3，Tra -1-60，Tra -1-81화전능형인자 C - MYC KLF -4 OCT -4 SOX -2，기태류조직절편중분별유소화선내피（내배층）、지방조직（중배층）화신경조직（외배층）등조직형성，설명해배태간세포형성적기태류유향삼배층분화적잠능。결론성공건립료인배태간세포건계，소건립적배양인배태간세포적방법가행。
Objective To investigate the method of human embryonic stem cells line,whether can provide a reliable cell research method and technology for study of stem cell and regenerative medicine field. Methods The feeder layer and no feeder layer system to culture human embryonic stem cells were applied respectively. And then immunofluorescence method,RT - PCR were used. These methods were used to identify its characteristic of stem cells of teratomas in vivo. Results The human embryonic stem cells express the stem cell surface marker, for example Oct - 4,SSEA - 3,Tra - 1 - 60,Tra - 1 - 81,and universal factors such as C - MYC KLF - 4 OCT - 4 SOX - 2,when culturing the human embryonic stem cells to thirty - three passages. The paraffin slice of teratomas can differentiate to digestive glands,fat tissue and nerve tissue,which originated from endoderm,mesoderm or ectodermal respectively. Therefore,the teratomas have the potential to differentiate to three germinal layer. Conclusion This result indicate that the human embryonic stem cells line is successful. The method of culturing the human stem cell is achievable.