title: Comparison of proliferative capacity of genetically-engineered pig and human corneal endothelial cells.
abstract: OBJECTIVE
  The possibility of providing cultured corneal endothelial cells (CECs) for clinical transplantation has gained much attention. However, the worldwide need for human (h) donor corneas far exceeds supply. The pig (p) might provide an alternative source. The aim of this study was to compare the proliferative capacity of CECs from wild-type (WT) pigs, genetically-engineered (GE) pigs, and humans.
  
  
  METHODS
  The following CECs were cultured: hCECs from donors (i) ≤36 years (young), (ii) ≥49 years (old), and WT pCECs from (iii) neonatal (<5 days), (iv) young (<2 months), and (v) old (>20 months) pigs, and CECs from young (vi) GE pigs (GTKO/CD46 and GTKO/CD46/CD55). Proliferative capacity of CECs was assessed by direct cell counting over 15 days of culture and by BrdU assay. Cell viability during culture was assessed by annexin V staining. The MTT assay assessed cell metabolic activity.
  
  
  RESULTS
  There was significantly lower proliferative capacity of old CECs than of young CECs (p < 0.01) in both pigs and humans. There was no significant difference in proliferative capacity/metabolic activity between young pCECs and young hCECs. However, there was a significantly higher percentage of cell death in hCECs compared to pCECs during culture (p < 0.01). Young GE pCECs showed similar proliferative capacity/cell viability/metabolic activity to young WT pCECs.
  
  
  CONCLUSIONS
  Because of the greater availability of young pigs and the excellent proliferative capacity of cultured GE pCECs, GE pigs could provide a source of CECs for clinical transplantation.
authors.affiliation: Department of Surgery, Thomas E. Starzl Transplantation Institute University of Pittsburgh, Pittsburgh, PA 15261, USA.
authors.forename: Minoru
authors.initials: M
authors.lastname: Fujita
authors.forename: Ruhina
authors.initials: R
authors.lastname: Mehra
authors.forename: Seung Eun
authors.initials: SE
authors.lastname: Lee
authors.forename: Danny S
authors.initials: DS
authors.lastname: Roh
authors.forename: Cassandra
authors.initials: C
authors.lastname: Long
authors.forename: James L
authors.initials: JL
authors.lastname: Funderburgh
authors.forename: David L
authors.initials: DL
authors.lastname: Ayares
authors.forename: David K C
authors.initials: DK
authors.lastname: Cooper
authors.forename: Hidetaka
authors.initials: H
authors.lastname: Hara
mesh_terms: D000328:Adult; D000818:Animals; D030801:Animals, Genetically Modified; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D004728:Endothelium, Corneal / Q000166:cytology*; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D013552:Swine / Q000235:genetics*; D013778:Tetrazolium Salts / Q000378:metabolism; D013844:Thiazoles / Q000378:metabolism
chemical_list: D013778:Tetrazolium Salts; D013844:Thiazoles; C022616:thiazolyl blue