Gagik Greg Haroutunian, Ashot Tsaghikian, Elena Fedorova, Pratima Chaurasia, Gabriele Luca Gusella, Arevik Mosoian
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Abstract
Rapid advances in mesenchymal stem progenitor cells (MSPCs) have rendered impetus into the area of cell therapy and regenerative medicine. The main promise of future stem cell therapies is their reliance on autologous stem cells derived from adipose tissue, which also includes treatments of bone fractures and degeneration. The effectiveness of different electric devices utilized to reprogram MSPCs toward osteogenic differentiation has provided varying degrees of effectiveness for clinical use. Adipose tissue-derived MSPCs were flow-cytometrically characterized and further differentiated into osteoblasts by culturing either in growth medium with pro-osteogenic supplements or without supplements with alternating electromagnetic field (EMF) generated by IteraCoil. IteraCoil is a multi-solenoid coil with a specific complex geometry that creates a 3D-EMF with desired parameters without directly applying electrodes to the cells and tissues. The flow-cytometric analysis of highly enriched (≥95%) adipose-derived MSPCs (CD34− , CD73+ , CD90+ , and CD105+ ) was utilized for the study. Osteoblasts and chondrocyte differentiations were then assessed by specific staining and quantified using ImageJ (National Institutes of Health). The osteoblastic differentiation of MSPCs cultured in regular medium and exposed to EMF at 0.05 and 1 kHz frequencies was compared with MSPCs cultured in a pro-osteogenic supplemented medium. In this study, we demonstrated that EMF from IteraCoil might have affected the signaling pathways that induce the osteogenic differentiation of human adipose-derived MSPCs in the absence of exogenous osteogenic factors. Therefore, EMF-generated osteogenic differentiation of reprogrammed adipose-derived autologous MSPCs may treat the loss of osteoblasts and osteoporosis and open new avenues for the development of regenerative cellular therapy. © 2022 Bioelectromagnetics Society.
利用IteraCoil装置产生的电磁场将间充质干细胞祖细胞分化成成骨谱系
间充质干细胞(MSPCs)的快速发展为细胞治疗和再生医学领域提供了动力。未来干细胞治疗的主要前景是依赖于来自脂肪组织的自体干细胞,这也包括骨折和退行性变的治疗。不同的电子设备用于MSPCs向成骨分化的重编程的有效性为临床应用提供了不同程度的有效性。用流式细胞术对脂肪组织来源的MSPCs进行了表征,并通过在含有促成骨补充剂或不含含IteraCoil产生的交变电磁场(EMF)的生长培养基中培养,进一步分化为成骨细胞。IteraCoil是一种多螺线管线圈,具有特定的复杂几何形状,可创建具有所需参数的3D-EMF,而无需直接将电极应用于细胞和组织。流式细胞术分析了高富集(≥95%)脂肪来源的MSPCs (CD34−,CD73+, CD90+和CD105+)。然后通过特异性染色评估成骨细胞和软骨细胞分化,并使用ImageJ(美国国立卫生研究院)进行定量。在常规培养基中培养的MSPCs和在0.05和1khz频率下暴露于EMF的MSPCs的成骨分化与在促成骨补充培养基中培养的MSPCs进行比较。在这项研究中,我们证明了在缺乏外源性成骨因子的情况下,来自IteraCoil的EMF可能会影响诱导人脂肪源性MSPCs成骨分化的信号通路。因此,emf诱导的重编程脂肪来源的自体MSPCs的成骨分化可能会治疗成骨细胞的缺失和骨质疏松症,并为再生细胞治疗的发展开辟新的途径。©2022生物电磁学学会。
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