Dynamic Three-Dimensional Culture Methods Enhance Mesenchymal Stem Cell Properties and Increase Therapeutic Potential
Dynamic Three-Dimensional Culture Methods Enhance Mesenchymal Stem Cell Properties and Increase Therapeutic Potential
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DOI：
10.1089/ten.tec.2009.0432
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发表时间：
2010-08-01
影响因子：
3
通讯作者：
Genever, Paul G.
中科院分区：
文献类型：
--
作者：
Frith, Jessica E.;Thomson, Brian;Genever, Paul G.
间充质干细胞（Mesenchymal stem cells，MSCs）具有自我更新和沿着分化成骨、成软骨和成脂的能力，在一系列治疗中具有潜在的应用价值。MSC可以在组织培养塑料上作为单层培养，但有迹象表明，它们在体外随着时间的推移而失去细胞特异性，因此不能很好地反映体内MSC的行为。我们开发了动态三维（3D）技术在体外MSC培养使用转瓶和旋转壁容器生物反应器。我们的特点是动态三维MSC培养的两种方法，并比较这些文化与单层MSC的属性。我们的研究结果表明，在最佳条件下，MSC形成紧凑的细胞球体，并在动态三维培养中保持活力。我们证明了改变细胞大小和表面抗原表达，以及增强成骨和成脂分化潜力的MSC从动态三维条件。通过对单层和转瓶MSC的微阵列分析，我们确定了基因表达的许多差异，包括那些证实细胞结构和细胞外基质广泛变化的差异。动态3D培养物中白细胞介素24的上调显示出选择性地损害在由动态3D MSC调节的培养基中培养的前列腺癌细胞的活力。总的来说，这项工作提出了一种新的治疗应用动态三维间充质干细胞，并证明这些方法是一个可行的替代单层技术，并可能证明有利于保留MSC的特性在体外。
Mesenchymal stemcells (MSCs) are capable of self-renewal and differentiation along the osteogenic, chondrogenic, and adipogenic lineages and have potential applications in a range of therapies. MSCs can be cultured as monolayers on tissue culture plastic, but there are indications that they lose cell-specific properties with time in vitro and so poorly reflect in vivo MSC behavior. We developed dynamic three-dimensional (3D) techniques for in vitro MSC culture using spinner flasks and a rotating wall vessel bioreactor. We characterized the two methods for dynamic 3D MSC culture and compared the properties of these cultures with monolayer MSCs. Our results showed that under optimal conditions, MSCs form compact cellular spheroids and remain viable in dynamic 3D culture. We demonstrated altered cell size and surface antigen expression together with enhanced osteogenic and adipogenic differentiation potential in MSCs from dynamic 3D conditions. By microarray analysis of monolayer and spinner flask MSCs, we identified many differences in gene expression, including those confirming widespread changes to the cellular architecture and extracellular matrix. The upregulation of interleukin 24 in dynamic 3D cultures was shown to selectively impair the viability of prostate cancer cells cultured in medium conditioned by dynamic 3D MSCs. Overall, this work suggests a novel therapeutic application for dynamic 3D MSCs and demonstrates that these methods are a viable alternative to monolayer techniques and may prove beneficial for retaining MSC properties in vitro.