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.
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Date
2014-02
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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).
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 × 106 cells) than Group I (0.04 × 106 cells). In Phase II, the average cell count was 5.44 × 104 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.
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Keywords
Corneal endothelium, human corneal endothelial precursor ells, in vitro expansion sphere, forming assay, thermo‑reversible gelation polymer, transportation
Citation
Rao Srinivas K, Sudhakar John, Parikumar Periyasamy, Natarajan Sundaram, Insaan Aditya, Yoshioka Hiroshi, Mori Yuichi, Tsukahara Shigeo, Baskar Subramani, Manjunath Sadananda Rao, Senthilkumar Rajappa, Thamaraikannan Paramasivam, Srinivasan Thangavelu, Preethy Senthilkumar, Abraham Samuel J K. 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. Indian Journal of Ophthalmology. 2014 Feb ; 62 (2): 130-135.