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脂肪来源干细胞条件培养基在聚己内酯纳米纤维上培养时可提高诱导多能干细胞的成骨分化。

Adipose-derived stem cells-conditioned medium improved osteogenic differentiation of induced pluripotent stem cells when grown on polycaprolactone nanofibers.

机构信息

Dietary Supplements and Probiotic Research Center, Alborz University of Medical Sciences, Karaj, Iran.

Urogenital Stem Cell Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

出版信息

J Cell Physiol. 2019 Jul;234(7):10315-10323. doi: 10.1002/jcp.27697. Epub 2018 Oct 30.

Abstract

Considering that the common osteogenic growth factors cannot be transplanted with stem cells to the patients, many studies are underway to find a replacement for these factors. Recently, it has been determined that mesenchymal stem cell (MSC)-derived conditioned medium (CM) contains effective factors in the bone formation process. In the current study, the synergistic effect of adipose-derived MSC's CM, and polycaprolactone (PCL) scaffold was investigated on the osteogenic differentiation potential of human induced pluripotent stem cells (iPSCs). After scaffold fabrication by electrospinning and characterization by scanning electron microscopy, iPSCs proliferation in the presence of CM, PCL, and both was evaluated using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide. Then, iPSCs osteogenic differentiation was investigated while cultured on tissue culture plate and PCL under CM compared with the osteogenic medium using alizarin red staining, calcium content, alkaline phosphatase activity and gene and protein expression analysis. Proliferation rate of the iPSCs was increased while cultured under CM and its effect was synergistically enhanced by culture on PCL. Evaluation of the osteogenic markers was showed CM alone could induce osteogenic differentiation into the iPSCs and this potential was significantly increased while combined with PCL nanofibrous scaffold. According to the results, it was demonstrated that CM has an osteogenic induction property almost the same of the common osteogenic medium and it can also be used potentially with stem cells when transplant to the patients. CM can also help by prolonging cell survival at the site of the defect as well as accelerating healing process.

摘要

考虑到常见的成骨生长因子不能与干细胞一起移植到患者体内,许多研究正在进行,以寻找这些因子的替代品。最近,已经确定间充质干细胞(MSC)衍生的条件培养基(CM)中含有骨形成过程中的有效因子。在本研究中,研究了脂肪来源的 MSC 的 CM 与聚己内酯(PCL)支架的协同作用对人诱导多能干细胞(iPSC)成骨分化潜能的影响。通过静电纺丝制备支架,并通过扫描电子显微镜进行表征后,用 3-[4,5-二甲基噻唑-2-基]-2,5 二苯基四氮唑溴盐评估 CM、PCL 和两者存在时 iPSC 的增殖情况。然后,通过茜素红染色、钙含量、碱性磷酸酶活性以及基因和蛋白表达分析,研究了 iPSC 在组织培养板和 CM 中培养时的成骨分化情况,与成骨培养基相比。iPSC 在 CM 中培养时增殖率增加,而在 PCL 上培养时协同作用增强。对成骨标志物的评估表明,CM 本身可以诱导 iPSC 向成骨细胞分化,而与 PCL 纳米纤维支架结合后,这种潜能显著增强。根据结果表明,CM 具有与普通成骨培养基几乎相同的成骨诱导特性,并且在移植到患者体内时也可以与干细胞一起使用。CM 还可以通过延长缺陷部位的细胞存活时间以及加速愈合过程来提供帮助。

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