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

• Published in: Indian journal of ophthalmology 2014-02, Vol.62 (2), p.130-135
• Main Authors: Rao, Srinivas, Sudhakar, John, Parikumar, Periyasamy, Natarajan, Sundaram, Insaan, Aditya, Yoshioka, Hiroshi, Mori, Yuichi, Tsukahara, Shigeo, Baskar, Subramani, Manjunath, Sadananda, Senthilkumar, Rajappa, Thamaraikannan, Paramasivam, Srinivasan, Thangavelu, Preethy, Senthilkumar, Abraham, Samuel
• Format: Article
• Language: English
• Subjects: Accommodation; Adolescent; Adult; Aged; Aged, 80 and over; Amblyopia; Blindness; Cadaver; cataract; Cell culture; citicoline; Cornea; Corneal endothelium; Corneal Transplantation; Endothelium; Endothelium, Corneal - cytology; Endothelium, Corneal - transplantation; Epidermal growth factor; Expansion; eye lens; human corneal endothelial precursor cells; Humans; in vitro expansion; In Vitro Techniques - methods; India; Low vision care; Middle Aged; multiple disabilities and visual impairment; Original; patching; Physiological aspects; Polymers; Polymers - pharmacology; presbyopia; Preservation; reading performance; risk factors; sphere forming assay; Stem cells; supraciliary segment implants; Temperature; thermo-reversible gelation polymer; Tissue Donors; Tissue Preservation - methods; Transplantation; Transplants & implants; transportation; Transportation - methods; treatment of cataract; Tropical Climate; Young Adult
• ISSN: 0301-4738; 1998-3689
• DOI: 10.4103/0301-4738.116457

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). Materials and 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. Conclusion: 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.