title: Successful transportation of human corneal endothelial tissues without cool preservation in varying Indian tropical climatic conditions and in vitro cell expansion using a novel polymer.
abstract: BACKGROUND
  Though the transplantation of human corneal endothelial tissue (CET) separated from cadaver cornea is in practice, its transportation has not been reported. We report the successful transportation of CET in varying Indian climatic conditions without cool preservation and the in vitro expansion of Human Corneal Endothelial Precursor Cells (HCEPCs) using a novel Thermo-reversible gelation polymer (TGP).
  
  
  METHODS
  CET from cadaver corneas (n = 67), unsuitable for transplantation, were used. In phase I, CET was transported in Basal Culture Medium (Group I) and TGP (Group II) and in Phase II, in TGP cocktail alone, from three hospitals 250-2500 km away, to a central laboratory. The transportation time ranged from 6 h to 72 h and the outdoor temperature between 20°C and 41°C. On arrival, CET were processed, cells were expanded upto 30 days in basal culture medium (Group A) and TGP scaffold (Group B). Cell viability and morphology were documented and Reverse transcription polymerase chain reaction (RT-PCR) characterization undertaken.
  
  
  RESULTS
  In Phase I, TGP yielded more viable cells (0.11 × 10(6) cells) than Group I (0.04 × 10(6) cells). In Phase II, the average cell count was 5.44 × 10(4) cells. During expansion, viability of HCEPCs spheres in TGP was maintained for a longer duration. The cells from both the groups tested positive for B-3 tubulin and negative for cytokeratins K3 and K12, thereby proving them to be HCEPCs.
  
  
  CONCLUSIONS
  TGP preserves the CET during transportation without cool preservation and supports in vitro expansion, with a higher yield of HCEPCs, similar to that reported in clinical studies.
authors.forename: Srinivas K
authors.initials: SK
authors.lastname: Rao
authors.forename: John
authors.initials: J
authors.lastname: Sudhakar
authors.forename: Periyasamy
authors.initials: P
authors.lastname: Parikumar
authors.forename: Sundaram
authors.initials: S
authors.lastname: Natarajan
authors.forename: Aditya
authors.initials: A
authors.lastname: Insaan
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Yoshioka
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Mori
authors.forename: Shigeo
authors.initials: S
authors.lastname: Tsukahara
authors.forename: Subramani
authors.initials: S
authors.lastname: Baskar
authors.forename: Sadananda Rao
authors.initials: SR
authors.lastname: Manjunath
authors.forename: Rajappa
authors.initials: R
authors.lastname: Senthilkumar
authors.forename: Paramasivam
authors.initials: P
authors.lastname: Thamaraikannan
authors.forename: Thangavelu
authors.initials: T
authors.lastname: Srinivasan
authors.forename: Senthilkumar
authors.initials: S
authors.lastname: Preethy
authors.affiliation: The Mary-Yoshio Translational Hexagon (MYTH), Nichi-In Centre for Regenerative Medicine, Chennai, India; II Department of Surgery, Yamanashi University - School of Medicine, Chuo, Japan, .
authors.forename: Samuel J K
authors.initials: SJ
authors.lastname: Abraham
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002102:Cadaver; D016039:Corneal Transplantation; D004728:Endothelium, Corneal / Q000166:cytology* / Q000637:transplantation; D006801:Humans; D066298:In Vitro Techniques / Q000379:methods; D007194:India; D008875:Middle Aged; D011108:Polymers / Q000494:pharmacology*; D014019:Tissue Donors; D014021:Tissue Preservation / Q000379:methods*; D014186:Transportation / Q000379:methods*; D014329:Tropical Climate; D055815:Young Adult
chemical_list: D011108:Polymers