cultivation of human corneal endothelial cells isolated from paired donor corneas培养的人类角膜内皮细胞分离配对捐献眼角膜.pdfVIP

Cultivation of Human Corneal Endothelial Cells Isolated from Paired Donor Corneas 1 1 1 1,2,4 1,2,3,4 Gary S. L. Peh , Kah-Peng Toh , Fei-Yi Wu , Donald T. Tan , Jodhbir S. Mehta * 1 Singapore Eye Research Institute, Singapore, Singapore, 2 Singapore National Eye Centre, Singapore, Singapore, 3 Duke Medical School of Medicine, National University of Singapore, Singapore, Singapore, 4 Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore Abstract Consistent expansion of human corneal endothelial cells (hCECs) is critical in the development of tissue engineered endothelial constructs. However, a wide range of complex culture media, developed from different basal media have been reported in the propagation of hCECs, some with more success than others. These results are further confounded by donor- to-donor variability. The aim of this study is to evaluate four culture media in the isolation and propagation of hCECs isolated from a series of paired donor corneas in order to negate donor variability. Isolated primary hCECs were cultured in four previously published medium coded in this study as: M1-DMEM; M2-OptiMEM-I; M3-DMEM/F12, M4-Ham’s F12/M199. Primary hCECs established in these conditions were expanded for two passages and analyzed for (1) their propensity to adhere and proliferate; (2) their expression of characteristic corneal endothelium markers: Na+K+/ATPase and ZO-1; and (3) their cellular morphology throughout the study. We found that hCECs isolated in all four media showed rapid attachment when cultured on FNC-coated dishes. However, hCECs established in the four media exhibited different proliferation profiles with striking morphological diffe