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R.J. Tsai; Ex vivo Expansion of Human Corneal Endothelial Cells for Clinical Application . Invest. Ophthalmol. Vis. Sci. 2006;47(13):2357.
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To develop techniques to culture the human corneal endothelial cells for clinical corneal endothelial keratoplasty.
Human donor corneas (from US Eye Bank) were selected for studies. Human corneal endothelium was obtained from the inner part of human limbal corneal rim. Endothelial sheet was cut into a small pieces and cultured on different substrata including amniotic membrane. Series passages were done. Cultured human corneal endothelial cells finally growth on the human corneal disc.
Age of the human donor corneas was from 37 y/o to 70 y/o with mean age of 63+11 y/o. The period of death to culture was from 6 days to 10 days (8.4 +1.8 days). The cultured human corneal endothelial cells (HCEC) could grow on the substrate with basement membrane substance. Serial passages from each HCEC were performed and results with 4.5 + 2.3 (2 to 9 passages) had been achieved from the aged human corneas. We took one of p2 HCEC and cultured on the human corneal disc. The HCEC was confluent growth on the human corneal disc after one month of culture. TEM demonstrated mosaic pattern of HCEC on the cornea disc.
The ex vivo expansion of human corneal endothelial cells systems have been established. These techniques will allow us to further study of clinical application or the engineering of cornea.
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