Yoshida Keiichiro, Obata Shuichi, Ono Michio, Esaki Michiyo, Maejima Tatsuo, Sawada Hajime
Department of Histology and Cell Biology, Yokohama City University School of Medicine, Fukuura 3-9, Kanazawa-ku, Yokohama, Kanagawa-ken 236-0004, Japan.
Eur J Cell Biol. 2007 Aug;86(8):461-71. doi: 10.1016/j.ejcb.2007.04.003. Epub 2007 Jun 27.
The 5F9A cell, which is a mesenchymal stem cell-like clone established from rat bone marrow substrate adherent cells, can differentiate into adipocytes and osteoblasts in vitro under the appropriate conditions. Multinucleated cells could be also induced by 12-O-tetradecanoylphorbol 13-acetate (TPA) in 5F9A cells. This effect was mediated by protein kinase C. Possible mechanisms of multinucleation by TPA were hypothesized to be either karyokinesis without cytokinesis or cell-cell fusion. By observation using time-lapse phase-contrast microscopy, we determined that the multinucleated cells were generated mainly by karyokinesis without cytokinesis. Cell fusion was studied using time-lapse photography, and confocal laser scanning microscopy using two differentially labeled cells. These techniques demonstrated that multinucleated 5F9A cells could be produced by cell fusion, albeit at a low frequency. We conclude that multinucleated 5F9A cells are formed primarily by karyokinesis without cytokinesis, although some cells are also formed by cell-cell fusion.
5F9A细胞是从大鼠骨髓基质贴壁细胞中建立的间充质干细胞样克隆，在适当条件下可在体外分化为脂肪细胞和成骨细胞。12 - O - 十四烷酰佛波醇13 - 乙酸酯（TPA）也可在5F9A细胞中诱导产生多核细胞。这种效应由蛋白激酶C介导。TPA诱导多核化的可能机制被推测为无胞质分裂的核分裂或细胞 - 细胞融合。通过延时相差显微镜观察，我们确定多核细胞主要是由无胞质分裂的核分裂产生的。使用延时摄影以及对两个用不同荧光标记的细胞进行共聚焦激光扫描显微镜观察来研究细胞融合。这些技术表明，尽管频率较低，但多核5F9A细胞可通过细胞融合产生。我们得出结论，多核5F9A细胞主要由无胞质分裂的核分裂形成，尽管也有一些细胞是由细胞 - 细胞融合形成的。
