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首页> 外文期刊>Biotechnology Progress >Accelerated adipogenic differentiation of hMSCs in a microfluidic shear stimulation platform
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Accelerated adipogenic differentiation of hMSCs in a microfluidic shear stimulation platform

机译：在微流剪切刺激平台中加速hMSCs的成脂分化

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The use of transplanted adipose tissue to repair crucial defects is clinically interesting for surgical reconstruction. Terminally differentiated adipocytes are utilized to promote the healthy regeneration of defective tissue. Use of differentiated mesenchymal stem cells, capable of differentiation into adipocytes, is advantageous because of their regenerative properties. Conventionally, the differentiation of hMSCs toward adipocytes occurs through chemical stimulation. We designed a microfluidic system, consisting of plastic tubing and a syringe pump, to create an environment of shear to accelerate this differentiation process. This system employed a flow rate equivalent to the accelerated flow rates found within the arterial system in order to promote and activate intracellular and extracellular proteins associated with the adipogenic lineage. Confirmation of sustained viability following shear exposure was obtained using a fluorescent live-dead assay. Visualization of intracellular lipid accumulation was achieved via Oil Red O staining. When placed into culture, shear stimulated hMSCs were further induced toward brown adipose tissue, as evidenced by a greater quantity of lipid triglycerides, relative to unstimulated hMSCs. qRT-PCR analysis validated the phenotypic changes observed when the hMSCs were later cultured in adipogenic differentiation media. Additionally, increased fold change for adipogenic markers such as LPL1, CFL1, and SSP1 were observed as a result of shear stimulation. The significance of this work lies in the demonstration that transient fluid shear exposure of hMSCs in suspension can influence differentiation into adipocytes. (c) 2015 American Institute of Chemical Engineers Biotechnol. Prog., 32:440-446, 2016
机译：在临床上，使用移植的脂肪组织修复重要的缺损对于外科手术重建是有趣的。最终分化的脂肪细胞被用于促进缺陷组织的健康再生。能够分化为脂肪细胞的分化的间充质干细胞的使用具有优势，因为它们具有再生特性。常规地，hMSC向脂肪细胞的分化是通过化学刺激发生的。我们设计了一个微流体系统，该系统由塑料管和注射泵组成，以创建剪切环境，以加速该分化过程。为了促进和激活与成脂谱系相关的细胞内和细胞外蛋白，该系统采用与动脉系统内的加速流速相当的流速。使用荧光活死试验获得了剪切暴露后持续生存力的确认。通过油红O染色实现细胞内脂质蓄积的可视化。当置于培养物中时，相对于未刺激的hMSC，大量的脂质甘油三酸酯证明了剪切刺激的hMSC进一步被诱导向棕色脂肪组织。 qRT-PCR分析验证了hMSCs随后在成脂分化培养基中培养时观察到的表型变化。此外，由于剪切刺激，还观察到了成脂标记物（例如LPL1，CFL1和SSP1）的倍数变化增加。这项工作的意义在于证明悬浮液中hMSCs的瞬时流体剪切暴露会影响向脂肪细胞的分化。 （c）2015美国化学工程师学会生物技术学会。 Prog。，32：440-446，2016

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