Human amniotic epithelial cells as novel feeder layers for promoting ex vivo expansion of limbal epithelial progenitor cells

Ying Ting Chen, Wei Li, Yasutaka Hayashida, Hua He, Szu Yu Chen, David Y. Tseng, Ahmad Kheirkhah, Scheffer C.G. Tseng

Research output: Contribution to journalArticlepeer-review

69 Scopus citations


Human amniotic epithelial cells (HAECs) are a unique embryonic cell source that potentially can be used as feeder layers for expanding different types of stem cells. In vivo, HAECs uniformly expressed pan-cytokeratins (pan-CK) and heterogeneously expressed vimentin (Vim). The two phenotypes expressing either pan-CK(+)/Vim(+) or pan-CK(+)/Vim(-) were maintained in serum-free media with high calcium. In contrast, all HAECs became pan-CK(+)/Vim(+) in serum-containing media, which also promoted HAEC proliferation for at least eight passages, especially supplemented with epidermal growth factor and insulin. Mitomycin C-arrested HAEC feeder layers were more effective in promoting clonal growth of human limbal epithelial progenitors than conventional 3T3 murine feeder layers. Cells in HAEC-supported clones were uniformly smaller, sustained more proliferation, and expressed less CK12 and connexin 43 but higher levels of stem cell-associated markers such as p63, Musashi-1, and ATP-binding cassette subfamily G2 than those of 3T3-supported clones. Subculturing of clonally expanded limbal progenitors from HAEC feeder layers, but not from 3T3 feeder layers, gave rise to uniformly p63-positive epithelial progenitor cells as well as nestin-positive neuronal-like progenitors. Collectively, these results indicated that HAECs can be used as a human feeder layer equivalent for more effective ex vivo expansion of adult epithelial stem cells from the human limbus.

Original languageEnglish (US)
Pages (from-to)1995-2005
Number of pages11
JournalStem Cells
Issue number8
StatePublished - Aug 2007
Externally publishedYes


  • ATP-binding cassette subfamily G2
  • Amniotic epithelial cells
  • Amniotic membrane
  • Connexin 43
  • Cytokeratins K12
  • Ex vivo expansion
  • Feeder layers
  • Limbus
  • Musashi-1
  • Neuronal progenitors
  • Plasticity
  • Progenitor cells
  • Stem cells
  • p63

ASJC Scopus subject areas

  • Medicine(all)


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