title: Establishment of an untransfected human corneal stromal cell line and its biocompatibility to acellular porcine corneal stroma.
abstract: OBJECTIVE
  To establish an untransfected human corneal stromal (HCS) cell line and characterize its biocompatibility to acellular porcine corneal stroma (aPCS).
  
  
  METHODS
  Primary culture was initiated with a pure population of HCS cells in DMEM/F12 media (pH 7.2) containing 20% fetal bovine serum and various necessary growth factors. The established cell line was characterized by growth property, chromosome analysis, tumorigenicity assay, expression of marker proteins and functional proteins. Furthermore, the biocompatibility of HCS cells with aPCS was examined through histological and immunocytochemistry analyses and with light, electron microscopies.
  
  
  RESULTS
  HCS cells proliferated to confluence 2 weeks later in primary culture and have been subcultured to passage 140 so far. A continuous untransfected HCS cell line with a population doubling time of 41.44 hours at passage 80 has been determined. Results of chromosome analysis, morphology, combined with the results of expression of marker protein and functional proteins suggested that the cells retained HCS cell properties. Furthermore, HCS cells have no tumorigenicity, and with excellent biocompatibility to aPCS.
  
  
  CONCLUSIONS
  An untransfected and non-tumorigenic HCS cell line has been established, and the cells maintained positive expression of marker proteins and functional proteins. The cell line, with excellent biocompatibility to aPCS, might be used for in vitro reconstruction of tissue-engineered HCS.
authors: Key Laboratory for Corneal Tissue Engineering, College of Marine Life Sciences, Ocean University of China, Qingdao 266003, Shandong Province, China.
authors: Ting-Jun
authors: TJ
authors: Fan
authors: Xiu-Zhong
authors: XZ
authors: Hu
authors: Jun
authors: J
authors: Zhao
authors: Ying
authors: Y
authors: Niu
authors: Wen-Zhuo
authors: WZ
authors: Zhao
authors: Miao-Miao
authors: MM
authors: Yu
authors: Yuan
authors: Y
authors: Ge