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Human Adipose-Derived Mesenchymal Stem Cells Cryopreservation and Thawing Decrease alpha 4-Integrin Expression

机译:人脂肪衍生的间充质干细胞冷冻保存和解冻减少α4-整联蛋白表达

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Aim. The effects of cryopreservation on adipose tissue-derived mesenchymal stem cells are not clearly documented, as there is a growing body of evidence about the importance of adipose-derived mesenchymal stem cells for regenerative therapies. The aim of this study was to analyze human adipose tissue-derived mesenchymal stem cells phenotypic expression (CD34, CD45, CD73, CD90, CD105, and CD49d), colony forming unit ability, viability, and differentiation potential before and after cryopreservation. Materials and Methods. 12 samples of the adipose tissue were collected from a healthy donor using the liposuction technique. The cell isolation was performed by enzymatic digestion and then the cells were cultured up to passage 2. Before and after cryopreservation the immunophenotype, cellular viability analysis by flow cytometer, colony forming units ability, differentiation potential into adipocytes and osteoblasts as demonstrated by Oil Red O and Alizarin Red staining, respectively. Results. The immunophenotypic markers expression was largely preserved, and their multipotency was maintained. However, after cryopreservation, the cells decreased alpha 4-integrin expression (CD49d), cell viability, and number of colony forming units. Conclusions. These findings suggest that ADMSC transplanted after cryopreservation might compromise the retention of transplanted cells in the host tissue. Therefore, further studies are warranted to standardize protocols related to cryopreservation to attain full benefits of stem cell therapy.
机译:目的。不清楚地记录冷冻保存对脂肪组织衍生的间充质干细胞的影响,因为存在关于脂肪衍生的间充质干细胞用于再生疗法的重要性的越来越多的证据。本研究的目的是分析人脂肪组织衍生的间充质干细胞表型表达(CD34,CD45,CD73,CD90,CD105和CD49D),在冷冻保存之前和之后的菌落形成单位能力,活力和分化潜力。材料和方法。使用脂肪技术从健康供体中收集12种脂肪组织的样品。细胞分离通过酶消化进行,然后将细胞培养至第2段。冷冻保存前后的免疫蛋白酶型,流式细胞仪,菌落形成单位能力,分化潜力进入脂肪细胞和成骨细胞,如油红色o所示。和茜素红染色。结果。免疫型标记物表达在很大程度上被保存,并保持了它们的多能率。然而,在冷冻保存之后,细胞减少了α4-整合蛋白表达(CD49D),细胞活力和菌落形成单元的数量。结论。这些发现表明,冷冻保存后移植可能会损害宿主组织中移植细胞的抑制。因此,需要进一步研究,以标准化与冷冻保存相关的协议,以获得干细胞疗法的全部益处。

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