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.
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.
复制标题
DOI：
10.4103/0301-4738.116457
复制
发表时间：
2014-02
影响因子：
3.1
通讯作者：
Abraham SJ
中科院分区：
文献类型：
--
作者：
Rao SK;Sudhakar J;Parikumar P;Natarajan S;Insaan A;Yoshioka H;Mori Y;Tsukahara S;Baskar S;Manjunath SR;Senthilkumar R;Thamaraikannan P;Srinivasan T;Preethy S;Abraham SJ
虽然从尸体角膜分离的人角膜内皮组织（CET）的移植已在实践中，但其运输尚未见报道。我们报告了 CET 在印度不同气候条件下的成功运输，无需冷藏，以及使用新型热可逆凝胶聚合物 (TGP) 体外扩增人角膜内皮前体细胞 (HCEPC)。使用来自尸体角膜 (n = 67) 的 CET，不适合移植。在第一阶段，CET 在基础培养基（第一组）和 TGP（第二组）中运输，在第二阶段，仅以 TGP 混合物从 250-2500 公里外的三家医院运输到中心实验室。运输时间为6小时至72小时，室外温度为20℃至41℃。到达后，进行 CET 处理，细胞在基础培养基（A 组）和 TGP 支架（B 组）中扩增长达 30 天。记录细胞活力和形态，并进行逆转录聚合酶链反应（RT-PCR）表征。在第一阶段，TGP 产生的活细胞（0.11 × 106 个细胞）比 I 组（0.04 × 106 个细胞）更多。在第二阶段，平均细胞计数为 5.44 × 104 个细胞。在扩张过程中，TGP 中 HCEPC 球体的活力得以维持较长时间。两组细胞的 B-3 微管蛋白检测呈阳性，细胞角蛋白 K3 和 K12 呈阴性，从而证明它们是 HCEPC。 TGP 在运输过程中保留 CET，无需冷藏，并支持体外扩增，HCEPC 产量更高，与临床研究报告相似。
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). 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. 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. 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.