Department of Biochemistry and Molecular Dentistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan.
Advanced Research Center for Oral and Craniofacial Sciences, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan.
Methods Mol Biol. 2023;2582:255-268. doi: 10.1007/978-1-0716-2744-0_17.
Although two-dimensional (2D) cultures from bone lineage cells are often used, it is well-known that this culture system is completely different from the in vivo bone matrix environment. In this paper, we describe a 3D culture method using both the mouse osteocytic cell line, MLO-Y4, and an osteocyte-enriched population of the cells isolated from mice. These cells are embedded in collagen gel with recombinant cellular communication network (CCN) factor proteins; then, osteoblasts or osteoclasts are inoculated and cultured on the collagen gel. Because this method mimics the in vitro bone matrix environment, it is useful for understanding the detailed mechanism of actions of CCN proteins in the bone matrix.
虽然骨系细胞的二维（2D）培养物经常被使用，但众所周知，这种培养系统与体内骨基质环境完全不同。在本文中，我们描述了一种使用小鼠骨原代细胞和从小鼠分离的富含骨原代细胞的 3D 培养方法。这些细胞被嵌入含有重组细胞通讯网络（CCN）因子蛋白的胶原凝胶中；然后，在胶原凝胶上接种和培养成骨细胞或破骨细胞。由于这种方法模拟了体外骨基质环境，因此对于理解 CCN 蛋白在骨基质中的详细作用机制非常有用。
