| Objective: To study the method of culturing limbal stem cells of rabbit in vitro, to investigate the treatment of rabbit eyes with limbal stem cell deficiency by transplanting cultured limbal stem cells on amniotic membrane.Methods: Limbal stem cells (LSC) tissues of rabbit were cultured in vitro and then the cells were cultured on acellular human amniotic membrane (AM) by using air-lifting culture. The growth of cells was observed and the cultured cells were identified by hematoxylin-eosin (HE) staining and AE1 immunohistochemical staining. The rabbit-eye models with limbal stem cell deficiency in the left eyes were made form 30 healthy New Zealand white rabbits by burned the cornea with severe alkali. Then the plants of cultured rabbit limbal stem cells on human amniotic membrane were transplanted to the burned eyes. The rabbits were divided into 3 groups randomly: the cultured limbal stem cells on amniotic membrane transplantation group (AM+LSC), the acellular amniotic membrane transplantation group (AM) and the control group. The corneal changes were observed by a slit lamp every week, and the corneal HE and AE1 immunohistochemical stain were made at 1 month.Results: The cultivated rabbit limbal stem cells grew well in vitro, and continued to differentiate, proliferate and form 3-4 multiple cell layers on acellular amniotic membrane, and the cells were attached to the underlying amniotic membrane firmly. The... |
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