Radial-Flow Bioreactor Enables Uniform Proliferation of Human Mesenchymal Stem Cells Throughout a Three-Dimensional Scaffold
Radial-Flow Bioreactor Enables Uniform Proliferation of Human Mesenchymal Stem Cells Throughout a Three-Dimensional Scaffold
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DOI：
10.1089/ten.tec.2011.0722
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发表时间：
2013-02-01
影响因子：
3
通讯作者：
Yoshinari, Masao
中科院分区：
文献类型：
--
作者：
Katayama, Aiko;Arano, Taichi;Yoshinari, Masao
从人骨髓中获得的间充质干细胞（MSCs）具有多能性，在体外已扩增分化为多种间胚层组织。为了创造用于植入的生物人工组织和器官，必须诱导这些细胞的增殖。本研究采用径向流生物反应器（RFB）在大型支架上诱导人间充质干细胞（hMSCs）的三维（3D）扩增。研究了这种膨胀对细胞特性的影响。为了制备预培养片材，首先将hMSCs接种到1型胶原片材上，孵育12小时，然后将其放置在RFB中制备支架。以3mL/min的速度循环培养基，在37℃下动态培养1周，作为对照，也在培养皿中进行静态培养。分析了细胞的膨胀和特性。在成骨诱导培养基中动态培养后，研究了hMSCs中碱性磷酸酶（ALP）的活性，以探索其成骨分化的潜力。在动态培养1周时，与静态条件下相比，观察到细胞数量增加了约60%，并且在整个支架中分布均匀；hMSC标记物未见变化。在RFB中培养后，hMSCs仍保持成骨分化的能力。目前的结果表明，在RFB中3D动态培养可以使hMSCs均匀扩增，而细胞特性没有变化，这表明该技术在组织工程中的实用性。
Mesenchymal stem cells (MSCs) obtained from human bone marrow are pluripotent and have been expanded and differentiated into several kinds of mesodermal tissue in vitro. To create bioartificial tissues and organs for implantation, it is necessary to induce proliferation in such cells. In this study, a radial-flow bioreactor (RFB) was used to induce three-dimensional (3D) expansion of human MSCs (hMSCs) on a large scaffold. The effect of this expansion on cellular characteristics was investigated. To produce precultured sheets, the hMSCs were first seeded onto type 1 collagen sheets and incubated for 12 h, after which they were placed in the RFB for fabrication of scaffolds. The culture medium was circulated at 3mL/min, and the cells were dynamically cultured for 1 week at 37 degrees C. As a control, static cultivation in a culture dish was also carried out. Cellular expansion and characteristics were analyzed. Alkaline phosphatase (ALP) activity in the hMSCs was also investigated after dynamic culture in an osteogenesis induction medium to explore their potential for osteogenic differentiation. At 1 week of dynamic cultivation, a > 60% increase was observed in a number of cells together with a uniform distribution throughout the scaffolds compared with under static conditions; no change in hMSC markers was observed. The hMSCs retained the ability for osteogenic differentiation after culture in the RFB. The present results indicate that 3D dynamic culture in an RFB enables uniform expansion of hMSCs with no change in cellular characteristics, suggesting the usefulness of this technique in tissue engineering.