In vitro models of angiogenesis are valuable tools for understanding the underlying mechanisms of pathological conditions and for the preclinical evaluation of therapies. Our laboratory developed the rat mesentery culture model as a new tool for investigating mechanistic cell–cell interactions at specific locations across intact blood and lymphatic microvascular networks ex vivo. The objective of this study was to report a method for evaluating the effect of aging on human stem cell differentiation into pericytes during angiogenesis in cultured microvascular networks. DiI labeled exogenous stem cells were seeded onto harvested adult Wistar rat mesenteric tissues and cultured in alpha-MEM + 1% serum for up to 5 days according to four experimental groups: (1) adult human adipose–derived stem cells (hASCs), (2) aged hASCs, (3) adult human bone marrow-derived stem cells (hBMSCs), and (4) aged hBMSCs. Angiogenesis per experimental group was supported by observation of increased vessel density and capillary sprouting. For each tissue per experimental group, a subset of cells was observed in typical pericyte location wrapped along blood vessels. Stem cell differentiation into pericytes was supported by the adoption of elongated pericyte morphology along endothelial cells and positive NG2 labeling. The percentage of cells in pericyte locations was not significantly different across the experimental groups, suggesting that aged mesenchymal stem cells are able to retain their differentiation capacity. Our results showcase an application of the rat mesentery culture model for aging research and the evaluation of stem cell fate within intact microvascular networks.

中文翻译：

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一种新颖的组织培养模型，用于评估衰老对成人微血管网络中干细胞命运的影响。


血管生成的体外模型是了解病理状况的潜在机制和临床前治疗评估的宝贵工具。我们的实验室开发了大鼠肠系膜培养模型，作为一种新工具，用于研究离体完整血液和淋巴微血管网络特定位置的机械细胞间相互作用。本研究的目的是报告一种评估衰老对培养微血管网络中血管生成过程中人类干细胞分化为周细胞的影响的方法。根据四个实验组，将 DiI 标记的外源干细胞接种到收获的成年 Wistar 大鼠肠系膜组织上，并在 alpha-MEM + 1% 血清中培养长达 5 天：(1) 成人脂肪干细胞 (hASC)、( 2) 老化的 hASC，(3) 成人骨髓源性干细胞 (hBMSC)，以及 (4) 老化的 hBMSC。血管密度增加和毛细血管萌芽的观察结果支持了每个实验组的血管生成。对于每个实验组的每个组织，在沿着血管包裹的典型周细胞位置观察到细胞子集。沿内皮细胞采用细长的周细胞形态和阳性 NG2 标记支持干细胞分化为周细胞。各实验组中周细胞位置的细胞百分比没有显着差异，这表明老化的间充质干细胞能够保留其分化能力。我们的结果展示了大鼠肠系膜培养模型在衰老研究和完整微血管网络内干细胞命运评估中的应用。