Centre for Ocular Regeneration, Prof. Brien Holden Eye Research Centre, Sudhakar and Sreekanth Ravi Stem Cell Biology Laboratory, Hyderabad Eye Research Foundation, L.V. Prasad Eye Institute, Hyderabad 500034, India.
Department of Biotechnology, National Institute of Technology, Warangal 506004, India.
Cells. 2021 May 19;10(5):1254. doi: 10.3390/cells10051254.
Corneal injuries are among the leading causes of blindness and vision impairment. Trauma, infectious keratitis, thermal and chemical (acids and alkali burn) injuries may lead to irreversible corneal scarring, neovascularization, conjunctivalization, and limbal stem cell deficiency. Bilateral blindness constitutes 12% of total global blindness and corneal transplantation remains a stand-alone treatment modality for the majority of end-stage corneal diseases. However, global shortage of donor corneas, the potential risk of graft rejection, and severe side effects arising from long-term use of immunosuppressive medications, demands alternative therapeutic approaches. Umbilical cord-derived mesenchymal stem cells can be isolated in large numbers using a relatively less invasive procedure. However, their role in injury induced corneal repair is largely unexplored. Here, we isolated, cultured and characterized mesenchymal stem cells from human umbilical cord, and studied the expression of mesenchymal (CD73, CD90, CD105, and CD34), ocular surface and epithelial (PAX6, WNT7A, and CK-8/18) lineage markers through immunofluorescence. The cultured human limbal and corneal epithelial cells were used as controls. Scratch assay was used to study the corneal epithelial repair potential of umbilical cord-derived mesenchymal stem cells, in vitro. The in vitro cultured umbilical cord-derived mesenchymal stem cells were plastic adherent, showed trilineage differentiation and expressed: mesenchymal markers CD90, CD105, CD73; epithelial marker CK-8/18, and ocular lineage developmental markers PAX6 and WNT-7A. Our findings suggest that umbilical cord-derived mesenchymal stem cells promote repair of the injured corneal epithelium by stimulating the proliferation of corneal epithelial cells, in vitro. They may serve as a potential non-ocular source of stem cells for treating injury induced bilateral corneal diseases.
角膜损伤是导致失明和视力损害的主要原因之一。创伤、感染性角膜炎、热和化学(酸和碱烧伤)损伤可导致不可逆转的角膜瘢痕、新生血管形成、结膜化和角膜缘干细胞缺乏。双眼失明占全球总失明人数的 12%,角膜移植仍然是大多数终末期角膜疾病的独立治疗方法。然而,全球供体角膜短缺、移植物排斥的潜在风险以及长期使用免疫抑制药物引起的严重副作用,都需要替代治疗方法。脐带间充质干细胞可以通过相对较少的侵入性程序大量分离。然而,它们在损伤诱导的角膜修复中的作用在很大程度上尚未得到探索。在这里,我们从人脐带中分离、培养和鉴定了间充质干细胞,并通过免疫荧光法研究了间充质(CD73、CD90、CD105 和 CD34)、眼表面和上皮(PAX6、WNT7A 和 CK-8/18)谱系标志物的表达。培养的人角膜缘和角膜上皮细胞用作对照。划痕实验用于研究脐带间充质干细胞体外对角膜上皮修复的潜能。体外培养的脐带间充质干细胞为塑料贴壁细胞,具有三系分化能力,并表达:间充质标志物 CD90、CD105、CD73;上皮标志物 CK-8/18,以及眼部谱系发育标志物 PAX6 和 WNT-7A。我们的研究结果表明,脐带间充质干细胞通过刺激角膜上皮细胞的增殖,在体外促进受损角膜上皮的修复。它们可能成为治疗损伤诱导的双侧角膜疾病的非眼部来源的潜在干细胞。