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首页> 外文期刊>Acta biomaterialia >A bioengineering approach to Schlemm's canal-like stem cell differentiation for in vitro glaucoma drug screening
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A bioengineering approach to Schlemm's canal-like stem cell differentiation for in vitro glaucoma drug screening

机译：体外青光眼药物筛选施力施流性干细胞分化的生物工程方法

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摘要

Human Schlemm's canal (HSC) cells are critical for understanding outflow physiology and glaucoma etiology. However, primary donor cells frequently used in research are difficult to isolate. HSC cells exhibit both vascular and lymphatic markers. Human adipose-derived stem cells (ADSCs) represent a potential source of HSC due to their capacity to differentiate into both vascular and lymphatic endothelial cells, via VEGF-A and VEGF-C. Shear stress plays a critical role in maintaining HSC integrity, function, and PROX1 expression. Additionally, the human trabecular meshwork (HTM) microenvironment could provide cues for HSC-like differentiation. We hypothesize that subjecting ADSCs to VEGF-A or VEGF-C, shear stress, and co-culture with HTM cells could provide biological, mechanical, and cellular cues necessary for HSC-like differentiation. To test this hypothesis, effects of VEGF-A, VEGF-C, and shear stress on ADSC differentiation were examined and compared to primary HSC cells in terms of cell morphology, and HSC marker expression using qPCR, immunoblotting, and immunocytochemistry analysis. Furthermore, the effect of co-culture with HTM cells on porous scaffolds on ADSC differentiation was studied. Treatment with VEGF-C under shear stress is effective in differentiating ADSCs into PROX1-expressing HSC-like cells. Co-culture with HTM cells on porous scaffolds leads to HTM/ADSC-derived HSC-like constructs that regulate through-flow and respond as expected to dexamethasone.
机译：人类的施莱姆的运河（HSC）细胞对于了解流出生理和青光眼病因至关重要。然而，在研究中经常使用的主要供体细胞难以分离。 HSC细胞表现出血管和淋巴标记物。人类脂肪衍生的干细胞（ADSCs）代表了HSC的潜在来源，因为它们通过VEGF-A和VEGF-C分化成血管和淋巴内皮细胞。剪切应力在维护HSC完整性，功能和Prox1表达式中起着关键作用。另外，人的小梁网状（HTM）微环境可以提供用于HSC样分化的提示。我们假设对VEGF-A或VEGF-A或VEGF-C，剪切应力和与HTM细胞共同培养的影响可以提供HSC样分化所需的生物，机械和细胞提示。为了测试该假设，研究了VEGF-A，VEGF-C和剪切应力对ADSC分化的影响，并与使用QPCR，免疫印迹和免疫细胞化学分析的HSC标记表达的原发性HSC细胞进行比较。此外，研究了与HTM细胞对多孔支架上的ADSC分化对多孔支架的影响。用VEGF-C处理剪切应力处理是有效地将ADSC分化为表达的HSC样细胞。与多孔支架上的HTM细胞的共培养物导致HTM / ADSC衍生的HSC样构造，其调节流过流过，并按照预期对地塞米松进行响应。

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