Abstract
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.
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Acknowledgments
This work was supported in part by KAKENHI grants from the programs Grants-in-Aid for Scientific Research (C) to TN (#JP20K09889) and KAKENHI grants from the programs Grants-in-Aid for Scientific Research (B) to SK (#JP21H03105) and to MT (#JP19H03817) and Challenging Research (Pioneering) to MT (#JP20K20611) of the Japan Society for the Promotion of Science, Japan. We would like to thank to Ms. Yoshiko Miyake for secretarial assistance.
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Nishida, T., Kubota, S., Takigawa, M. (2023). Novel Cell Biological Assays for Measuring Bone Remodeling Activities of CCN Proteins. In: Takigawa, M. (eds) CCN Proteins. Methods in Molecular Biology, vol 2582. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2744-0_17
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DOI: https://doi.org/10.1007/978-1-0716-2744-0_17
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-2743-3
Online ISBN: 978-1-0716-2744-0
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