Sumi Keisuke, Abe Takaharu, Kunimatsu Ryo, Oki Nanae, Tsuka Yuji, Awada Tetsuya, Nakajima Kengo, Ando Kazuyo, Tanimoto Kotaro
Department of Orthodontics, Division of Oral Health and Development, Hiroshima University Hospital.
Department of Orthodontics and Craniofacial Developmental Biology, Hiroshima University Graduate School of Biomedical and Health Sciences.
J Oral Sci. 2018;60(2):221-225. doi: 10.2334/josnusd.17-0187.
Regeneration of tissue, including bone, using mesenchymal stem cells (MSCs) has been progressing rapidly. Regeneration of bone requires the presence of an appropriate environment and efficient chemotaxis of cells to the target site. Differentiation of MSCs into mesenchymal cells has received considerable attention, but the effect of MSCs on chemotaxis is not well understood. In this study, we investigated the effect of MSCs on chemotaxis of RAW264 cells via C-C motif chemokine ligand 2 (CCL2). Balb/c mouse bone marrow-derived MSCs and RAW264 cells, which are osteoclast precursor cells, were co-cultured without cell contact. The gene expression of CCL2 in MSCs and CC-chemokine receptor 2 (CCR2) in RAW264 cells was determined using quantitative real-time PCR. Analysis of RAW264 cell chemotaxis was performed using the Boyden chamber assay. mRNAs for CCL2 and CCR2 were significantly upregulated upon co-culture in comparison to culture of either cell type alone, and the number of chemotactic RAW264 cells was significantly increased by co-culture. MSCs enhanced the chemotaxis of RAW264 cells, possibly via CCL2-CCR2 interaction, suggesting the potential utility of MSCs for tissue regeneration.
利用间充质干细胞（MSC）进行包括骨组织在内的组织再生研究进展迅速。骨组织再生需要合适的环境以及细胞向靶位点的有效趋化作用。间充质干细胞向间充质细胞的分化已受到广泛关注，但间充质干细胞对趋化作用的影响尚不清楚。在本研究中，我们通过C-C基序趋化因子配体2（CCL2）研究了间充质干细胞对RAW264细胞趋化作用的影响。将Balb/c小鼠骨髓来源的间充质干细胞与作为破骨细胞前体细胞的RAW264细胞进行非接触共培养。使用定量实时PCR测定间充质干细胞中CCL2的基因表达以及RAW264细胞中CC趋化因子受体2（CCR2）的基因表达。使用Boyden小室法进行RAW264细胞趋化性分析。与单独培养任何一种细胞类型相比，共培养时CCL2和CCR2的mRNA显著上调，并且共培养显著增加了趋化性RAW264细胞的数量。间充质干细胞可能通过CCL2-CCR2相互作用增强RAW264细胞的趋化性，提示间充质干细胞在组织再生中的潜在应用价值。
