| 摘要 |
A method for expansion of limbal stem cells comprises: (i) providing a limbal biopsy on a human amniotic membrane; (ii) culturing for a first time period the limbal biopsy in the presence of epithelial culture medium comprising hydrocortisone, insulin, tri-iodothyronine, adenine, and epidermal growth factor, wherein the epithelial culture medium is supplemented with serum and one or more antibiotics; and (iii) substituting the epithelial culture medium of step ii) with epithelial culture medium comprising hydrocortisone, insulin, tri-iodothyronine, adenine, and epidermal growth factor, wherein the epithelial culture medium is supplemented with serum and is free from antibiotics, and culturing for a second time period; wherein the expanded limbal stem cells form an epithelial sheet on the human amniotic membrane and wherein the serum is derived from the same donor as the limbal biopsy. A defined epithelial culture medium comprising hydrocortisone, insulin, tri-iodothyronine, adenine, and epidermal growth factor, and optionally cholera toxin is provided for use in such a method; the culture medium may be supplemented with penicillin and/or streptomycin for use in step (ii) of the method. |
