Differentiation of Rodent Bone Marrow Mesenchymal Stem Cells into Intervertebral Disc-like Cells Following Coculture with Rat Disc Tissue
Differentiation of Rodent Bone Marrow Mesenchymal Stem Cells into Intervertebral Disc-like Cells Following Coculture with Rat Disc Tissue
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DOI：
10.1089/ten.tea.2008.0458
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发表时间：
2009-09-01
影响因子：
4.1
通讯作者：
Diwan, Ashish D.
中科院分区：
文献类型：
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作者：
Wei, Aiqun;Chung, Sylvia A.;Diwan, Ashish D.
本研究旨在探讨大鼠骨髓间充质干细胞（mesenchymal stem cells，rMSCs）与椎间盘组织共培养能否诱导分化为椎间盘样细胞。将rMSC与啮齿动物椎间盘一起在transwell板中培养长达30天。采用免疫组化、Western blot、real-time RT-PCR、北方印迹、电镜等方法检测rMSCs的分化情况。并对细胞的多向分化潜能、蛋白多糖和胶原合成潜能进行了研究。共培养14天后，rMSCs发生形态学变化，形成三维微团，并在RNA和蛋白水平表达胶原蛋白-2，aggrecan和sox-9。而单独培养的rMSCs则未出现上述变化。共培养的rMSCs还表现出其他盘状细胞的特征，包括细胞外基质的形成，蛋白多糖和胶原蛋白的合成。此外，通过电子显微镜观察共培养的rMSCs与椎间盘组织之间的细胞接触。当局部环境改变时，定向的rMSCs仍保留其向脂肪细胞或骨细胞的中胚层谱系分化的能力。这项研究支持，骨髓间充质干细胞是一个有前途的来源，细胞治疗和组织工程在椎间盘再生，并强调，rMSCs可以诱导成髓核样细胞在体外的直接影响下，完整的椎间盘组织。
This study aimed to evaluate whether rat mesenchymal stem cells (rMSCs) could be differentiated in vitro into disc-like cells by coculturing with intervertebral disc tissue. rMSCs were cultured with rodent intervertebral disc for up to 30 days in transwell plates. The differentiation of rMSCs was evaluated by immunostaining, Western blot, real-time RT-PCR, Northern blot, and electron microscopy. The potentials of multilineage differentiation and proteoglycan and collagen synthesis were also investigated. rMSCs underwent morphological changes to form three-dimensional micromasses and expressed collagen-2, aggrecan, and sox-9 at RNA and protein levels after 14 days of coculture. These changes were not detected in the samples of rMSCs cultured alone. Cocultured rMSCs also showed other characteristic features of disc-like cells, including the extracellular matrix formation, and proteoglycan and collagen synthesis. In addition, cellular contact between cocultured rMSCs and disc tissue was observed by electron microscopy. Committed rMSCs still retained their differentiation ability into mesoderm lineages of adipocytes or osteocytes when the local environment was altered. This study supports that MSCs are a promising source for cell therapy and tissue engineering in disc regeneration, and highlights that rMSCs can be induced into nucleus pulposus-like cells in vitro under the direct influence of intact disc tissue.