This invention relates to a method for the reconstruction of a tissue-engineered human corneal endothelium. Human corneal endothelial cells are cultured in vitro to logarithmic growth phase using 20% calf bovine serum-containing DMEM/F12 medium. Trypsin is used to digest epithelial layer of the freeze-dried human amniotic membrane in order to produce denuded human amniotic membrane as scaffold carriers. The scaffold carriers are tiled on the bottom of culture plate wells until they are dried and completely adhered to the bottom of wells. Human corneal endothelial cells at logarithmic growth phase are re-suspended in DMEM/F12 medium containing type-IV collagen and 20% calf bovine serum. Human corneal endothelial cell suspension is subsequently inoculated to amniotic membrane scaffold carriers that are tiled on the bottom of wells in culture plate to launch in vitro culture as well as in vitro reconstruction of tissue-engineered human corneal endothelium. This invention is scientific and rational. The reconstructed tissue-engineered human corneal endothelium can be produced on large scale to satisfy the great demand of tissue-engineered human corneal endothelium in clinical cornea transplantation for primary corneal endotheliopathy therapy. Meanwhile, costs for in vitro reconstruction of tissue-engineered human corneal endothelium and clinical therapy are low.
