College of Veterinary Medicine, Northwest A&F University, Shaanxi Stem Cell Engineering and Technology Center, Yangling 712100, China.
Exp Eye Res. 2013 Oct;115:246-54. doi: 10.1016/j.exer.2013.07.024. Epub 2013 Aug 8.
Limbal epithelial stem cells are responsible for the self-renewal and replenishment of the corneal epithelium. Although it is possible to repair the ocular surface using limbal stem cell transplantation, the mechanisms behind this therapy are unclear. To investigate the distribution of surviving donor cells in a reconstructed corneal epithelium, we screened a Venus-labeled limbal stem cell strain in goats. Cells were cultivated on denuded human amniotic membrane for 21 days to produce Venus-labeled corneal epithelial sheets. The Venus-labeled corneal epithelial sheets were transplanted to goat models of limbal stem cell deficiency. At 3 months post-surgery, the damaged corneal epithelia were obviously improved in the transplanted group compared with the non-transplanted control, with the donor cells still residing in the reconstructed ocular surface epithelium. Using Venus as a marker, our results indicated that the location and survival of donor cells varied, depending on the corneal epithelial region. Additionally, immunofluorescent staining of the reconstructed corneal epithelium demonstrated that many P63(+) cells were unevenly distributed among basal and suprabasal epithelial layers. Our study provides a new model, and reveals some of the mechanisms involved in corneal epithelial cell regeneration research.
角膜缘上皮干细胞负责角膜上皮的自我更新和补充。虽然可以通过角膜缘干细胞移植来修复眼表面，但这种治疗的机制尚不清楚。为了研究供体细胞在重建的角膜上皮中的分布情况，我们筛选了山羊中标记有 Venus 的角膜缘干细胞系。将细胞在去表皮的人羊膜上培养 21 天，以产生 Venus 标记的角膜上皮片。将 Venus 标记的角膜上皮片移植到山羊角膜缘干细胞缺陷模型中。术后 3 个月，与未移植对照组相比，移植组受损的角膜上皮明显改善，供体细胞仍存在于重建的眼表面上皮中。使用 Venus 作为标记物，我们的结果表明，供体细胞的位置和存活取决于角膜上皮区域。此外，重建的角膜上皮的免疫荧光染色表明，许多 P63(+)细胞在基底和超基底上皮层中不均匀分布。我们的研究提供了一个新的模型，并揭示了一些角膜上皮细胞再生研究中涉及的机制。
