3D spheroid culture system on micropatterned substrates for improved differentiation efficiency of multipotent mesenchymal stem cells
3D spheroid culture system on micropatterned substrates for improved differentiation efficiency of multipotent mesenchymal stem cells
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DOI：
10.1016/j.biomaterials.2009.01.030
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发表时间：
2009-05-01
期刊：
影响因子：
14
通讯作者：
Kataoka, Kazunori
中科院分区：
文献类型：
--
作者：
Wang, Wenjie;Itaka, Keiji;Kataoka, Kazunori
多能间充质干细胞（MSCs）是再生医学中最强大的工具之一。然而，其较低的分化效率限制了MSCs在临床治疗中的进一步应用。在这里，我们报告使用基于光刻和微图像化技术的3D球体培养方法可以实现MSCs向成体细胞的更高多能分化效率。首先在微畴基质上制备尺寸精确、质量均匀的间充质干细胞球体，然后将其诱导成脂肪细胞和成骨细胞。RT-PCR的基因表达结果和形态学观察结果均表明，三维球体培养法可显著提高分化效率。基因芯片分析获得的基因表达谱证实了MSCs的高分化效率，并揭示了在3D球体培养系统中诱导的MSCs不仅通过增加脂肪生成和成骨相关基因的表达水平，还通过下调维持MSCs自我更新表型的基因来调节基因表达。我们认为，我们的三维球形培养系统有助于优化MSCs的高效分化，为研究工程化三维培养系统的高效分化机制提供了新的思路，并有望在再生医学和药物研发领域得到广泛应用。2009爱思唯尔有限公司版权所有。
Multipotent mesenchymal stem cells (MSCs) are one of the most powerful tools in regeneration medicine. Their low differentiation efficiency, however, limits further application of MSCs in clinical therapy. Here we report that a much higher multipotent differentiation efficiency of MSCs to adult cells can be achieved using a 3D spheroid culture method based on photolithography and micropatterning techniques. MSC spheroid of precise dimension and uniform quality cultured on the microdomain substrates was prepared first, and then was induced into adipocytes and osteoblasts. Both gene expression results from RT-PCR and morphology observation results revealed that the 3D spheroid culture method could greatly improve differentiation efficiency. Gene expression profiles obtained from gene microarray analysis confirmed the high differentiation efficiency and revealed that MSCs induced in 3D spheroid culture system regulated gene expression not only by increasing the expression levels of genes related to adipogenesis and osteogenesis, but also by down-regulating the gene maintaining MSCs' self-renewal phenotypes. We conclude that our 3D spheroid culture system contributes to an optimization for efficient differentiation of MSCs, offers insight into the mechanism of efficient differentiation of engineered 3D culture system, and has promise for wide applications in regeneration medicine and drug discovery fields. (C) 2009 Elsevier Ltd. All rights reserved.