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首页> 外文期刊>Molecular reproduction and development >Reconstruction of corneal epithelium with cryopreserved corneal limbal stem cells in a goat model.
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Reconstruction of corneal epithelium with cryopreserved corneal limbal stem cells in a goat model.

机译：在山羊模型中用冷冻保存的角膜缘干细胞重建角膜上皮。

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We describe a procedure to construct an artificial corneal epithelium from cryopreserved limbal stem cells (LSCs) for corneal transplantation. The LSCs were separated from limbal tissue of male goats. The primary LSCs were identified by flow cytometry and were expanded. They were examined for stem cell-relevant properties and cryopreserved in liquid nitrogen. Cryopreserved LSCs were thawed and then transplanted onto human amniotic membrane, framed on a nitrocellulose sheet, to construct corneal epithelium sheets. The artificial corneal epithelium was transplanted into the right eye of pathological models of total limbal stem cell deficiency (LSCD). Then, the effects of reconstruction were evaluated by clinical observation and histological examination. Polymerase chain reaction analysis was used to detect the SRY gene. The data showed that transplantation of cryopreserved LSCs, like fresh LSCs, successfully reconstructed damaged goat corneal surface gradually, but the SRY gene expression from male goat cells could only be detected in the first 2 months after transplantation. The therapeutic effect of the transplantation may be associated with the inhibition of inflammation-related angiogenesis after transplantation of cryopreserved LSCs. This study provides the first line of evidence that cryopreserved LSCs can be used for reconstruction of damaged corneas, presenting a remarkable potential source for transplantation in the treatment of corneal disorders.
机译：我们描述了从冷冻保存的角膜缘干细胞（LSCs）中构建人工角膜上皮用于角膜移植的程序。从雄山羊的角膜缘组织分离LSC。通过流式细胞术鉴定了主要的LSC，并进行了扩增。检查了它们的干细胞相关特性，并将其冷冻保存在液氮中。将冷冻保存的LSC解冻，然后移植到人羊膜上，并置于硝酸纤维素片上，以构建角膜上皮片。将人工角膜上皮移植到全角膜缘干细胞缺乏症（LSCD）病理模型的右眼中。然后，通过临床观察和组织学检查评估重建的效果。聚合酶链反应分析用于检测SRY基因。数据显示，冷冻保存的LSCs像新鲜LSCs一样，可以成功地逐步重建受损的山羊角膜表面，但是雄性山羊细胞的SRY基因表达只能在移植后的前两个月检测到。移植的治疗作用可能与冷冻保存的LSC移植后抑制炎症相关的血管生成有关。这项研究提供的第一线证据是，冷冻保存的LSC可以用于重建受损的角膜，为角膜疾病的治疗提供了重要的潜在移植来源。

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