Gurel Mediha, Zomer Helena, McFetridge Calum, Murfee Walter L, McFetridge Peter S
J. Crayton Pruitt Family Department of Biomedical Engineering, University of Florida, Gainesville, Florida, USA.
Biotechnology Research and Application Center, Cukurova University, Adana, Turkey.
Microcirculation. 2025 Apr;32(3):e70007. doi: 10.1111/micc.70007.
Mesenchymal stem cells (MSCs) represent an attractive option as an endothelial cell (EC) source for regenerative medicine therapies. However, the differentiation of MSCs toward an ECs phenotype can be regulated by a complex and dynamic microenvironment, including specific growth factors as well as local mechanical cues. The objective of this work was to evaluate whether Physiologically-modeled dynamic stimulation (PMDS) characterized by continuous variability in pulse frequencies mimicking the dynamic temporal range of cardiac function would enhance MSC differentiation toward ECs compared to a constant frequency stimulation.
Mesenchymal stem cells were grown in a complex growth factor cocktail versus standard culture media to initiate the endothelial differentiation process, then subsequently exposed to PMDS that vary in duration and constant flow (CF) at a fixed 10 dynes/cm shear stress and 1.3 Hz frequency.
Both PMDS and media type strongly influence cell differentiation and function. Cells were shown to significantly upregulate eNOS activity and displayed lower TNF-a induced leukocyte adhesion compared to cells cultured under CF, consistent with a more quiescent ECs phenotype that regulates anti-inflammatory and anti-thrombotic states.
These findings suggest that the dynamic microenvironment created by perfusion, in contrast to constant frequency, combined with growth factors, enhances MSCs differentiation toward a vascular endothelial-like phenotype.
间充质干细胞（MSCs）作为再生医学疗法中内皮细胞（ECs）的来源具有吸引力。然而，MSCs向内皮细胞表型的分化可受复杂且动态的微环境调节，包括特定生长因子以及局部机械信号。本研究的目的是评估以模拟心脏功能动态时间范围的脉冲频率持续变化为特征的生理模型动态刺激（PMDS）与恒定频率刺激相比，是否能增强MSCs向内皮细胞的分化。
间充质干细胞在复合生长因子混合物中与标准培养基中培养以启动内皮分化过程，随后暴露于在固定的10达因/平方厘米剪切应力和1.3赫兹频率下持续时间不同的PMDS及恒定流（CF）。
PMDS和培养基类型均强烈影响细胞分化和功能。与在CF条件下培养的细胞相比，这些细胞显示出eNOS活性显著上调，且TNF-α诱导的白细胞黏附较低，这与调节抗炎和抗血栓状态的更静止的内皮细胞表型一致。
这些发现表明，与恒定频率相比，由灌注与生长因子结合所产生的动态微环境可增强MSCs向血管内皮样表型的分化。
