Immunophenotypical Characterization of Limbal Mesenchymal Stromal Cell Subsets during In Vitro Expansion

IF 5.4 3区材料科学 Q2 CHEMISTRY, PHYSICAL

ACS Applied Energy Materials Pub Date : 2024-08-09 DOI:10.3390/ijms25168684

Sara Aghazadeh, Q. Peng, F. Dardmeh, Jesper Hjortdal, Vladimir Zachar, H. Alipour

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Abstract

Limbal mesenchymal stromal cells (LMSCs) reside in the limbal niche, supporting corneal integrity and facilitating regeneration. While mesenchymal stem/stromal cells (MSCs) are used in regenerative therapies, there is limited knowledge about LMSC subpopulations and their characteristics. This study characterized human LMSC subpopulations through the flow cytometric assessment of fifteen cell surface markers, including MSC, wound healing, immune regulation, ASC, endothelial, and differentiation markers. Primary LMSCs were established from remnant human corneal transplant specimens and passaged eight times to observe changes during subculture. The results showed the consistent expression of typical MSC markers and distinct subpopulations with the passage-dependent expression of wound healing, immune regulation, and differentiation markers. High CD166 and CD248 expressions indicated a crucial role in ocular surface repair. CD29 expression suggested an immunoregulatory role. Comparable pigment-epithelial-derived factor (PEDF) expression supported anti-inflammatory and anti-angiogenic roles. Sustained CD201 expression indicated maintained differentiation capability, while VEGFR2 expression suggested potential endothelial differentiation. LMSCs showed higher VEGF expression than fibroblasts and endothelial cells, suggesting a potential contribution to ocular surface regeneration through the modulation of angiogenesis and inflammation. These findings highlight the heterogeneity and multipotent potential of LMSC subpopulations during in vitro expansion, informing the development of standardized protocols for regenerative therapies and improving treatments for ocular surface disorders.

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体外扩增过程中眼睑间充质基质细胞亚群的免疫表型特征描述

角膜缘间充质基质细胞（LMSCs）存在于角膜缘龛中，支持角膜完整性并促进角膜再生。虽然间充质干细胞/基质细胞（MSCs）被用于再生疗法，但人们对LMSC亚群及其特征的了解还很有限。本研究通过对15种细胞表面标志物（包括间充质干细胞、伤口愈合、免疫调节、ASC、内皮细胞和分化标志物）进行流式细胞术评估，确定了人类LMSC亚群的特征。原代 LMSCs 从残留的人类角膜移植标本中提取，并经过八次传代培养，以观察亚培养过程中的变化。结果显示，典型的间充质干细胞标志物和不同的亚群表达一致，伤口愈合、免疫调节和分化标志物的表达与传代有关。CD166 和 CD248 的高表达表明它们在眼表修复中起着关键作用。CD29 的表达表明其具有免疫调节作用。类似的色素上皮衍生因子（PEDF）表达支持抗炎和抗血管生成的作用。CD201 的持续表达表明其具有维持分化的能力，而 VEGFR2 的表达则表明其具有潜在的内皮分化能力。与成纤维细胞和内皮细胞相比，LMSCs 的血管内皮生长因子表达量更高，这表明它们可通过调节血管生成和炎症为眼表再生做出潜在贡献。这些发现凸显了 LMSC 亚群在体外扩增过程中的异质性和多潜能性，为制定再生疗法的标准化方案和改善眼表疾病的治疗提供了参考。

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来源期刊

ACS Applied Energy Materials Materials Science-Materials Chemistry

CiteScore

10.30

自引率

6.20%

发文量

1368

期刊介绍： ACS Applied Energy Materials is an interdisciplinary journal publishing original research covering all aspects of materials, engineering, chemistry, physics and biology relevant to energy conversion and storage. The journal is devoted to reports of new and original experimental and theoretical research of an applied nature that integrate knowledge in the areas of materials, engineering, physics, bioscience, and chemistry into important energy applications.