caa a22 4500 605462909 CHVBK 20210128100250.0 cr unu---uuuuu 210128e20150101xx s 000 0 eng 10.1007/s00774-013-0555-5 doi (NATIONALLICENCE)springer-10.1007/s00774-013-0555-5 Establishment of optimized in vitro assay methods for evaluating osteocyte functions [Elektronische Daten] [Masashi Honma, Yuki Ikebuchi, Yoshiaki Kariya, Hiroshi Suzuki] Recent studies have revealed that osteocytes play multiple important physiological roles. To analyze osteocyte functions in detail, an in vitro experimental system for primary osteocytes would be useful. Unfortunately, osteocytes tend to dedifferentiate and acquire osteoblast-like features even when the cells are cultured in three-dimensional (3D) collagen gel. Therefore, it is desirable to establish osteocyte culture conditions that prevent dedifferentiation over longer periods. In this study, we obtained systematic information about the influence of culture conditions on osteocyte differentiation states. Fetal bovine serum (FBS) concentrations from 0.1 to 0.5% in 3D culture matrix did not significantly influence the expression of osteocyte markers. On the other hand, addition of Matrigel to the culture matrix significantly enhanced the expression of Rankl and late osteocyte markers such as Sost and Fgf23. Matrigel addition also inhibited upregulation of Opg and early osteocyte markers such as Dmp1 and Gp38. These effects on osteocyte properties were maximal at a Matrigel culture matrix content of 50%. Matrigel addition to the matrix also increased dendritic process extension by osteocytes. In addition, Matrigel addition significantly stimulated tartrate-resistant acid phosphatase activity in co-culture with bone marrow macrophages. Among the conditions tested, 50% Matrigel and 0.2% FBS in type I collagen matrix were optimal for culture of primary osteocytes. The Japanese Society for Bone and Mineral Research and Springer Japan, 2013 Osteocyte nationallicence Differentiation nationallicence Culture method nationallicence Osteoclastogenesis nationallicence Co-culture system nationallicence Honma Masashi Department of Pharmacy, The University of Tokyo Hospital, Faculty of Medicine, The University of Tokyo, 7-3-1 Hongo, 113-8655, Bunkyo-ku, Tokyo, Japan aut Ikebuchi Yuki Department of Pharmacy, The University of Tokyo Hospital, Faculty of Medicine, The University of Tokyo, 7-3-1 Hongo, 113-8655, Bunkyo-ku, Tokyo, Japan aut Kariya Yoshiaki Department of Pharmacy, The University of Tokyo Hospital, Faculty of Medicine, The University of Tokyo, 7-3-1 Hongo, 113-8655, Bunkyo-ku, Tokyo, Japan aut Suzuki Hiroshi Department of Pharmacy, The University of Tokyo Hospital, Faculty of Medicine, The University of Tokyo, 7-3-1 Hongo, 113-8655, Bunkyo-ku, Tokyo, Japan aut Journal of Bone and Mineral Metabolism Springer Japan 33/1(2015-01-01), 73-84 0914-8779 33:1<73 2015 33 774 https://doi.org/10.1007/s00774-013-0555-5 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00774-013-0555-5 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Honma Masashi Department of Pharmacy, The University of Tokyo Hospital, Faculty of Medicine, The University of Tokyo, 7-3-1 Hongo, 113-8655, Bunkyo-ku, Tokyo, Japan aut NATIONALLICENCE 700 1- Ikebuchi Yuki Department of Pharmacy, The University of Tokyo Hospital, Faculty of Medicine, The University of Tokyo, 7-3-1 Hongo, 113-8655, Bunkyo-ku, Tokyo, Japan aut NATIONALLICENCE 700 1- Kariya Yoshiaki Department of Pharmacy, The University of Tokyo Hospital, Faculty of Medicine, The University of Tokyo, 7-3-1 Hongo, 113-8655, Bunkyo-ku, Tokyo, Japan aut NATIONALLICENCE 700 1- Suzuki Hiroshi Department of Pharmacy, The University of Tokyo Hospital, Faculty of Medicine, The University of Tokyo, 7-3-1 Hongo, 113-8655, Bunkyo-ku, Tokyo, Japan aut NATIONALLICENCE 773 0- Journal of Bone and Mineral Metabolism Springer Japan 33/1(2015-01-01), 73-84 0914-8779 33:1<73 2015 33 774