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唇黏膜干细胞：分离、鉴定及其在角膜上皮重建中的潜在应用。

Labial Mucosa Stem Cells: Isolation, Characterization, and Their Potential for Corneal Epithelial Reconstruction.

机构信息

Institute of Cytology Russian Academy of Science, St. Petersburg, Russia.

Department of Cytology and Histology, St. Petersburg State University, St. Petersburg, Russia.

出版信息

Invest Ophthalmol Vis Sci. 2022 Jul 8;63(8):16. doi: 10.1167/iovs.63.8.16.

DOI:10.1167/iovs.63.8.16
PMID:35848889
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9308017/
Abstract

PURPOSE

The purpose of this study was to characterize labial mucosa stem cells (LMSCs) and to investigate their potential for corneal epithelial reconstruction in a rabbit model of total limbal stem cell deficiency (LSCD).

METHODS

Rabbit LMSCs (rLMSCs) and human (hLMSCs) LMSCs were derived from labial mucosa and characterized in terms of their proliferation activity by the evaluation of proliferation index (PI) and colony forming efficiency (CFE), cell senescence, and differentiation abilities. The expression of various limbus-specific, stem cell-specific, and epithelial markers was assessed via immunocytochemistry. Flow cytometry was used to evaluate mesenchymal and hematopoietic cell surface markers expression. Chromosomal stability of the derived cells was examined using the conventional GTG-banding technique. To assess the impact of LMSCs on corneal epithelial reconstruction, rLMSCs were seeded onto a decellularized human amniotic membrane (dHAM), thereafter their regeneration potential was examined in the rabbit model of total LSCD.

RESULTS

Both rLMSCs and hLMSCs showed high proliferation and differentiation abilities, entered senescence at later passages, and expressed different stem cell-specific (ABCB5, ALDH3A1, ABCG2, and p63α), mesenchymal (vimentin), and epithelial (CK3/12, CK15) markers. Cell surface antigen expression was similar to other described mesenchymal stem cells. No clonal structural chromosome abnormalities (CSCAs) and the low percentage of non-clonal structural chromosome abnormalities (NSCAs) were observed. Transplantation of rLMSCs promoted corneal epithelial reconstruction and enhanced corneal transparency.

CONCLUSIONS

LMSCs have significant proliferation and differentiation abilities, display no detrimental chromosome aberrations, and demonstrate considerable potential for corneal repair.

摘要

目的

本研究旨在对唇黏膜干细胞（LMSCs）进行特征分析，并在兔全角膜缘干细胞缺失（LSCD）模型中研究其对角膜上皮重建的潜力。

方法

从唇黏膜中分离并鉴定兔 LMSCs（rLMSCs）和人 LMSCs（hLMSCs），通过增殖指数（PI）和集落形成效率（CFE）评估来评估增殖活性，检测细胞衰老和分化能力。通过免疫细胞化学评估各种角膜缘特异性、干细胞特异性和上皮标记物的表达。采用流式细胞术评估间充质和造血细胞表面标记物的表达。采用常规 GTG 带技术检测细胞的染色体稳定性。为了评估 LMSCs 对角膜上皮重建的影响，将 rLMSCs 接种到脱细胞人羊膜（dHAM）上，然后在兔全 LSCD 模型中检测其再生潜能。

结果

rLMSCs 和 hLMSCs 均显示出较高的增殖和分化能力，在后期传代时进入衰老状态，并表达不同的干细胞特异性（ABCB5、ALDH3A1、ABCG2 和 p63α）、间充质（波形蛋白）和上皮（CK3/12、CK15）标记物。细胞表面抗原表达与其他描述的间充质干细胞相似。未观察到克隆结构染色体异常（CSCAs）和低比例的非克隆结构染色体异常（NSCAs）。rLMSCs 移植促进了角膜上皮的重建，增强了角膜的透明度。

结论

LMSCs 具有显著的增殖和分化能力，无不良染色体畸变，具有很大的角膜修复潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/928c/9308017/eb46cb9edfca/iovs-63-8-16-f008.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/928c/9308017/9248a79da72f/iovs-63-8-16-f005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/928c/9308017/af197df0edce/iovs-63-8-16-f006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/928c/9308017/8851755c0bf5/iovs-63-8-16-f007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/928c/9308017/eb46cb9edfca/iovs-63-8-16-f008.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/928c/9308017/68cfa4598042/iovs-63-8-16-f002.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/928c/9308017/0fe3b3a270c2/iovs-63-8-16-f004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/928c/9308017/9248a79da72f/iovs-63-8-16-f005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/928c/9308017/af197df0edce/iovs-63-8-16-f006.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/928c/9308017/eb46cb9edfca/iovs-63-8-16-f008.jpg

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