title: Utilization of human amniotic mesenchymal cells as feeder layers to sustain propagation of human embryonic stem cells in the undifferentiated state.
abstract: 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.
authors: Department of Immunology, Institute of Clinical Medicine, China-Japan Friendship Hospital, Beijing, People's Republic of China.
authors: Kehua
authors: K
authors: Zhang
authors: Zhe
authors: Z
authors: Cai
authors: Yang
authors: Y
authors: Li
authors: Jun
authors: J
authors: Shu
authors: Lin
authors: L
authors: Pan
authors: Fang
authors: F
authors: Wan
authors: Hong
authors: H
authors: Li
authors: Xiaojie
authors: X
authors: Huang
authors: Chun
authors: C
authors: He
authors: Yanqiu
authors: Y
authors: Liu
authors: Xiaohui
authors: X
authors: Cui
authors: Yang
authors: Y
authors: Xu
authors: Yan
authors: Y
authors: Gao
authors: Liqun
authors: L
authors: Wu
authors: Shanxia
authors: S
authors: Cao
authors: Lingsong
authors: L
authors: Li
mesh_terms: D000650:Amnion / Q000166:cytology*; D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D018920:Coculture Techniques; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000502:physiology; D061252:Feeder Cells / Q000166:cytology*; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000502:physiology; D051379:Mice; D013724:Teratoma / Q000473:pathology