稳定表达E-cadherin的TW2R细胞增强人胚胎干细胞生长  

E-cadherin-expressing TW2R feed cells enhance growth of undifferentiated human embryonic stem cells

作　　者：宋琼 滕夏虹[3] 王丽惠 许倩倩 孙晓婷 邹春林 SONG Qiong;TENG Xiahong;WANG Lihui;XU Qianqian;SUN Xiaoting;ZOU Chunlin(Center for Translational Medicine and School of Preclinical Medicine,Guangxi Medical University,Nanning 530021,China;Key Laboratory of Longevity and Ageing-Related Disease of Chinese Ministry of Education,Nanning 530021;School of International Education,Guangxi Medical University,Nanning 530021)

机构地区：[1]广西医科大学转化医学研究中心,广西医科大学基础医学院,南宁530021 [2]长寿与老年相关疾病教育部重点实验室,南宁530021 [3]广西医科大学国际教育学院,南宁530021

出　　处：《中国比较医学杂志》2021年第11期1-8,共8页Chinese Journal of Comparative Medicine

基　　金：国家自然科学基金项目(81971191,81670750);教育部留学回国人员科研启动基金资助项目(教外司留[2013]694号)。

摘　　要：目的构建一种可高效支持人胚胎干细胞生长的人源饲养层细胞系。方法首先通过在TW2R细胞系稳定表达人E-钙黏素(E-cadherin)基因构建永生化人源饲养层细胞系TWE3R,并比较人胚胎干细胞在TWE3R及其亲本细胞上的克隆形成数。将H9人胚胎干细胞在TWE3R细胞上经过10代连续传代培养后,进行人胚胎干细胞特异性标志物分析、拟胚体形成分析和畸胎瘤形成检测等实验。结果本研究成功构建了永生化的人源饲养层细胞系TWE3R,TWE3R细胞可比其亲本细胞更高效地支持人胚胎干细胞生长,并且TWE3R饲养层细胞还具有支持低密度接种条件下(每平方厘米5个)人胚胎干细胞生长的特性。此外,H9人胚胎干细胞在TWE3R细胞上长期传代培养后,仍可保持所有人胚胎干细胞的未分化特性、正常细胞核型以及在体内外均可分化形成所有3个胚层衍生物的潜能。结论本研究所建立的永生化人源饲养层细胞系TWE3R具有高效支持胚胎干细胞生长的特性。Objective To establish an immortalized human feeder cell line that efficiently supports the growth of undifferentiated human embryonic stem(hES) cells. Methods The immortalized TWE3 R cell line was established by stably expressing the human E-cadherin gene in the TW2 R cell line. The numbers of hESC colonies on TWE3 R feeder cells were compared with those on its parental cell lines. After H9 hES cells were cultured on TWE3 R cells for 10 consecutive passages, pluripotency marker analysis, embryoid body(EB) formation assays and teratoma formation assays were conducted. Results We succeeded in establishing the TWE3 R human feeder cell line that showed higher efficiency to support the growth of hES cells than its parental cell line. TWE3 R feeder cells supported the growth of H9 ESCs at low density(5 cells/cm;). Additionally, after long-term culture on TWE3 R feeders, H9 ESCs retained all undifferentiated characteristics, a normal karyotype and the potential to differentiate into derivatives of all three embryonic germ layers. Conclusions The immortalized TWE3 R human feeder cell line is capable of supporting efficient growth of human ESCs.

关 键 词：人胚胎干细胞 人源饲养层细胞 E-钙黏素 

分 类 号：R-33[医药卫生]