Autologous transplantation of conjunctival epithelial stem cells

我们分别培养供眼和兔眼角膜和结膜上皮细胞。此外，我们开发了一种用于角膜和结膜上皮细胞的成体前体细胞/干细胞分离方法（球体形成试验）。我们证明，与培养的分化上皮细胞相比，BrDU染色显示角膜和结膜前体细胞/干细胞具有更高的增殖能力，巢蛋白染色显示未成熟状态。我们在羊膜上培养结膜上皮细胞，发现培养的结膜上皮在羊膜上形成3 ~ 4层。将培养的结膜上皮片移植到角膜上皮混浊、结膜浸润的兔角膜上后，移植兔角膜保持角膜清晰度。这些结果表明，利用培养的结膜上皮细胞将重建的结膜上皮片移植到角膜是治疗全角膜缘干细胞缺乏症的一种可能的治疗策略。

We cultured corneal and conjunctival epithelial cells from donor eye and rabbit eye. Additionally, we developed an adult precursor/stem cell isolation method(the sphere-forming assay) for corneal and conjunctival epithelial cells. We demonstrated corneal and conjunctival precursor/stem cells possessed the higher proliferative capacity as indicated by BrDU staining and the immature state as indicated by nestin staining compared with cultured differentiated epithelial cells. We cultured conjunctival epithelial cells on the denuded amniotic membrane and demonstrated that cultured conjuncdtival epithelium formed three to four layers on amniotic membrane. After transplantation of cultured conjunctival epithelial sheet on the rabbit cornea with corneal epithelial opacity and conjunctival invasion onto cornea, transplanted rabbit cornea maintained corneal clarity. These results suggest that transplantation of reconstructed conjunctival epithelial sheet using cultured conjunctival epithelial cells to the cornea is a possible treatment strategy for treating total limbal stem cell deficiency.