Clumps of Mesenchymal Stem Cell/Extracellular Matrix Complexes Generated with Xeno-Free Conditions Facilitate Bone Regeneration via Direct and Indirect Osteogenesis
Clumps of Mesenchymal Stem Cell/Extracellular Matrix Complexes Generated with Xeno-Free Conditions Facilitate Bone Regeneration via Direct and Indirect Osteogenesis
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DOI：
10.3390/ijms20163970
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发表时间：
2019-08-02
影响因子：
5.6
通讯作者：
Kurihara, Hidemi
中科院分区：
文献类型：
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作者：
Motoike, Souta;Kajiya, Mikihito;Kurihara, Hidemi
间充质干细胞（MSC）/细胞外基质（ECM）复合物（C-MSC）的三维团块由细胞和自身产生的ECM组成。我们之前已经证明，C-MSC可以在没有人工支架的情况下移植到骨缺损区域以诱导骨再生。为了将C-MSCs作为一种可靠的骨再生疗法应用于临床，本研究旨在在无异种/无血清条件下产生C-MSCs，其可以发挥成功的骨再生特性，并监测骨愈合过程中移植细胞和宿主细胞之间的相互作用。人骨髓源性MSC在无异种/无血清培养基中培养。为了获得C-MSC，使用微量移液管尖端刮擦在细胞片上形成的汇合细胞，然后撕下。将片材卷起以形成圆形细胞团。然后，将C-MSCs移植到免疫缺陷小鼠颅骨缺损模型中。C-MSCs的移植以时间依赖的方式诱导骨再生。免疫荧光染色显示，供体人细胞和宿主小鼠细胞都有助于骨重建。脱细胞C-MSCs移植后不能诱导骨再生，尽管宿主细胞可以浸润到缺损区。这些发现表明，在无异种/无血清条件下产生的C-MSCs可以通过直接和间接成骨诱导骨再生。
Three-dimensional clumps of mesenchymal stem cell (MSC)/extracellular matrix (ECM) complexes (C-MSCs) consist of cells and self-produced ECM. We demonstrated previously that C-MSCs can be transplanted into bone defect regions with no artificial scaffold to induce bone regeneration. To apply C-MSCs in a clinical setting as a reliable bone regenerative therapy, the present study aimed to generate C-MSCs in xeno-free/serum-free conditions that can exert successful bone regenerative properties and to monitor interactions between grafted cells and host cells during bone healing processes. Human bone marrow-derived MSCs were cultured in xeno-free/serum-free medium. To obtain C-MSCs, confluent cells that had formed on the cellular sheet were scratched using a micropipette tip and then torn off. The sheet was rolled to make a round clump of cells. Then, C-MSCs were transplanted into an immunodeficient mouse calvarial defect model. Transplantation of C-MSCs induced bone regeneration in a time-dependent manner. Immunofluorescence staining showed that both donor human cells and host mice cells contributed to bone reconstruction. Decellularized C-MSCs implantation failed to induce bone regeneration, even though the host mice cells can infiltrate into the defect area. These findings suggested that C-MSCs generated in xeno-free/serum-free conditions can induce bone regeneration via direct and indirect osteogenesis.