Utilization of human amniotic mesenchymal cells as feeder layers to sustain propagation of human embryonic stem cells in the undifferentiated state.
Utilization of human amniotic mesenchymal cells as feeder layers to sustain propagation of human embryonic stem cells in the undifferentiated state.
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DOI：
10.1089/cell.2010.0103
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发表时间：
2011-07
影响因子：
1.6
通讯作者：
Kehua Zhang;Zhe Cai;Yang Li;J. Shu;Lin Pan;Fang Wan;Hong Li;Xiao-jie Huang;C. He;Yanqiu Liu;Xiaohui Cui;Yang Xu;Yan Gao;Liqun Wu;Shanxia Cao;Ling-song Li
中科院分区：
文献类型：
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作者：
Kehua Zhang;Zhe Cai;Yang Li;J. Shu;Lin Pan;Fang Wan;Hong Li;Xiao-jie Huang;C. He;Yanqiu Liu;Xiaohui Cui;Yang Xu;Yan Gao;Liqun Wu;Shanxia Cao;Ling-song Li
人类胚胎干细胞(ES)通常通过在小鼠胚胎成纤维细胞(MEF)饲养层上培养来维持未分化状态。然而，MEF不适合支持临床使用的人类ES细胞，因为动物细胞有患人畜共患病的风险。因此，需要为人类ES细胞开发临床所需的人组织基饲养层。在此，我们报道了人羊膜间充质细胞(HAMCs)可以作为人ES细胞的饲养细胞，因为它们容易获得，并且相对不受伦理问题的影响。像MEF一样，hAMCs可以作为人类ES细胞生长良好的饲养细胞。集落直径和生长曲线测定及细胞周期分析表明，人ES细胞在HAMCs饲养层上的自我更新率高于MEF和人羊膜上皮细胞。免疫荧光染色和免疫印迹显示，人ES细胞在HAMCs上培养后表达干细胞标记物Oct-3/4、Sox2和NANOG。通过体外类胚体形成和体内畸胎瘤形成的验证，我们发现在HAMCs饲养层上培养的人ES细胞经过20代培养后仍能保持分化为三层胚层的能力。综上所述，我们的数据提示HAMCs可能是安全的饲养细胞，以支持处于未分化状态的人ES细胞的生长，用于未来的治疗。
Human embryonic stem (ES) cells are usually maintained in the undifferentiated state by culturing on feeder cells layers of mouse embryonic fibroblasts (MEFs). However, MEFs are not suitable to support human ES cells used for clinical purpose because of risk of zoonosis from animal cells. Therefore, human tissue-based feeder layers need to be developed for human ES cells for clinical purpose. Hereof we report that human amniotic mesenchymal cells (hAMCs) could act as feeder cells for human ES cells, because they are easily obtained and relatively exempt from ethical problem. Like MEFs, hAMCs could act as feeder cells for human ES cells to grow well on. The self-renewal rate of human ES cells cultured on hAMCs feeders was higher than that on MEFs and human amniotic epithelial cells determined by measurement of colonial diameters and growth curve as well as cell cycle analysis. Both immunofluorescence staining and immunoblotting showed that human ES cells cultured on hAMCs expressed stem cell markers such as Oct-3/4, Sox2, and NANOG. Verified by embryoid body formation in vitro and teratoma formation in vivo, we found out that after 20 passages of culture, human ES cells grown on hAMCs feeders could still retain the potency of differentiating into three germ layers. Taken together, our data suggested hAMCs may be safe feeder cells to sustain the propagation of human ES cells in undifferentiated state for future therapeutic use.