title: Stemness characteristics of human corneal endothelial cells cultured in various media.
abstract: OBJECTIVE
  To investigate stemness characteristics of human corneal endothelial cells (HCECs) cultured in various media.
  
  
  METHODS
  Human corneal endothelial cells were isolated using a sphere-forming assay. Cells were allowed to attach to the bottom of culture plates and were cultured in different media designated as medium A (Opti-MEM I with 8% fetal bovine serum), medium B (DMEM/F12 with B27 supplement), medium E (DMEM/F12 with epidermal growth factor [EGF]), and medium BE (DMEM/F12 with B27 supplement and EGF), respectively. Cell morphology was evaluated with an phase-contrast inverted microscope. Immunofluorescence staining and western blotting of nestin, octamer-binding transcription factor (OCT3/4), glial fibrillary acidic protein (GFAP), zonula occludens-1 (ZO-1), collagen VIII alpha2, and Na-K ATPase was performed. Cell proliferation was assessed with a cell counting kit-8 assay.
  
  
  RESULTS
  A few cultured cells stained with nestin. The cells cultured in medium A expressed high levels of GFAP, OCT3/4, and nestin, and higher levels of ZO-1 were expressed in the cells cultured in medium A and medium B compared with cells cultured in the other media. The cells cultured in medium A assumed a fibroblast-like shape, whereas the cells cultured in medium B and medium BE appeared as mosaics. Cell proliferation was highest in medium A compared with those cultured in the other media.
  
  
  CONCLUSIONS
  Cultured HCECs expressed stem cell markers, including nestin, OCT3/4, and GFAP. The expression of stem cell markers differed according to the culture media and associated proliferation rate.
authors.affiliation: Department of Ophthalmology (J.W.N., K.Y., Y.J.S.), Hallym University College of Medicine, Seoul, Korea; Department of Ophthalmology (J.J.K., J.Y.H.), Seoul National University Bundang Hospital, Seongnam, Korea; Department of Ophthalmology (J.Y.H., W.R.W.), Seoul National University College of Medicine, Seoul, Korea; and Department of Ophthalmology (E.-S.C., T.-Y.C.), Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.
authors.forename: Jin Woo
authors.initials: JW
authors.lastname: Noh
authors.forename: Jin Joo
authors.initials: JJ
authors.lastname: Kim
authors.forename: Joon Young
authors.initials: JY
authors.lastname: Hyon
authors.forename: Eui-Sang
authors.initials: ES
authors.lastname: Chung
authors.forename: Tae-Young
authors.initials: TY
authors.lastname: Chung
authors.forename: Kayoung
authors.initials: K
authors.lastname: Yi
authors.forename: Won Ryang
authors.initials: WR
authors.lastname: Wee
authors.forename: Young Joo
authors.initials: YJ
authors.lastname: Shin
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media / Q000494:pharmacology*; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D004728:Endothelium, Corneal / Q000166:cytology* / Q000187:drug effects; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D006801:Humans; D064231:Nestin / Q000378:metabolism; D050810:Octamer Transcription Factors / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*
chemical_list: D015415:Biomarkers; D003470:Culture Media; D005904:Glial Fibrillary Acidic Protein; D064231:Nestin; D050810:Octamer Transcription Factors