Skin-Derived Precursors as a Source of Progenitors for Corneal Endothelial Regeneration.
Skin-Derived Precursors as a Source of Progenitors for Corneal Endothelial Regeneration.
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DOI：
10.1002/sctm.16-0162
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发表时间：
2017-03
影响因子：
6
通讯作者：
Shimmura S
中科院分区：
文献类型：
--
作者：
Inagaki E;Hatou S;Higa K;Yoshida S;Shibata S;Okano H;Tsubota K;Shimmura S
角膜失明是世界上第四大失明原因。目前的治疗方法是同种异体角膜移植，但受到供体短缺和免疫排斥的限制。皮肤源性前体细胞 (SKP) 是出生后干细胞，是自我更新的多能前体细胞，可以从真皮中分离和扩增。因此，面部皮肤可能是神经嵴衍生细胞的可利用的自体来源。从 Wnt1-Cre/Floxed EGFP 小鼠的面部皮肤中分离出 SKP。用含有视黄酸和 GSK 3-β 抑制剂的培养基诱导分化后，SKP 形成多边形角膜内皮样细胞 (sTECE)。通过逆转录聚合酶链反应（RT-PCR）和定量实时聚合酶链反应（qRT-PCR）证实了主要角膜内皮标志物的表达。蛋白质印迹证实了 Na,K-ATP 酶蛋白的表达，这是角膜内皮细胞的主要功能标志物。免疫组织化学揭示了细胞-细胞连接中闭锁小带-1 和 Na、K-ATP 酶的表达。 Na,K-ATP酶泵活性的体外功能分析表明，与 SKP 或对照 3T3 细胞相比，sTECE 具有显着较高的泵功能。此外，将sTECE移植到兔大疱性角膜病模型中，成功维持了角膜厚度和透明度。此外，我们成功地从人 SKP 中诱导出角膜内皮样细胞，并表明移植的角膜也保持了角膜透明度和厚度。我们的研究结果表明，SKP 可作为自体细胞的来源，用于治疗角膜内皮疾病。干细胞转化医学 2017；6：788–798
Corneal blindness is the fourth leading cause of blindness in the world. Current treatment is allogenic corneal transplantation, which is limited by shortage of donors and immunological rejection. Skin‐derived precursors (SKPs) are postnatal stem cells, which are self‐renewing, multipotent precursors that can be isolated and expanded from the dermis. Facial skin may therefore be an accessible autologous source of neural crest derived cells. SKPs were isolated from facial skin of Wnt1‐Cre/Floxed EGFP mouse. After inducing differentiation with medium containing retinoic acid and GSK 3‐β inhibitor, SKPs formed polygonal corneal endothelial‐like cells (sTECE). Expression of major corneal endothelial markers were confirmed by Reverse transcription polymerase chain reaction (RT‐PCR) and quantitative Real time polymerase chain reaction (qRT‐PCR). Western blots confirmed the expression of Na, K‐ATPase protein, the major functional marker of corneal endothelial cells. Immunohistochemistry revealed the expression of zonular occludens‐1 and Na, K‐ATPase in cell‐cell junctions. In vitro functional analysis of Na, K‐ATPase pump activity revealed that sTECE had significantly high pump function compared to SKPs or control 3T3 cells. Moreover, sTECE transplanted into a rabbit model of bullous keratopathy successfully maintained corneal thickness and transparency. Furthermore, we successfully induced corneal endothelial‐like cells from human SKPs, and showed that transplanted corneas also maintained corneal transparency and thickness. Our findings suggest that SKPs may be used as a source of autologous cells for the treatment of corneal endothelial disease. Stem Cells Translational Medicine 2017;6:788–798