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Cited by 500

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Establishment of an Osteocyte-like Cell Line, MLO-Y4

Kato, Yoichi, Windle, Jolene J., Koop, Barbara A., Mundy, Gregory R., Bonewald, Lynda F.·1997·Journal of Bone and Mineral Research
DOI 10.1359/jbmr.1997.12.12.2014
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Abstract

Although osteocytes are the most abundant cells in bone, their functional role remains unclear. In part, this is due to lack of availability of osteocyte cell lines which can be studied in vitro. Since others have shown that cell lines can be readily developed from transgenic mice in which the SV40 large T-antigen oncogene is expressed under the control of a promoter which targets the cells of interest, we used this approach to develop an osteocyte cell line. We chose as a promoter osteocalcin, whose expression is essentially limited to bone cells and which is expressed more abundantly in osteocytes than in osteoblasts. From these transgenic mice, we isolated cells from the long bones using sequential collagenase digestion and maintained these cells on collagen-coated surfaces which are optimal for osteocyte maintenance and growth. We describe here the properties of a cell line cloned from these cultures, called MLO-Y4 (for murine long bone osteocyte Y4). The properties of MLO-Y4 cells are very similar to primary osteocytes. Like primary osteocytes and unlike primary osteoblasts, the cell line produces large amounts of osteocalcin but low amounts of alkaline-phosphatase. The cells produce extensive, complex dendritic processes and are positive for T-antigen, for osteopontin, for the neural antigen CD44, and for connexin 43, a protein found in gap junctions. This cell line also produces very small amounts of type I collagen mRNA compared with primary osteoblasts. MLO-Y4 cells lack detectable mRNA for osteoblast-specific factor 2, which appears to be a positive marker for osteoblasts but may be a negative marker for osteocytes. This newly established cell line should prove useful for studying the effects of mechanical stress on osteocyte function and for determining the means whereby osteocytes communicate with other bone cells such as osteoblasts and osteoclasts.

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Cited by

Metadata

Authors
Kato, Y., Windle, J. J., Koop, B. A., Mundy, G. R., & Bonewald, L. F.
Affiliations
  • Department of Medicine, University of Texas Health Science Center at San Antonio, San Antonio, Texas, U.S.A.
  • Cancer Therapy and Research Center, University of Texas Health Science Center at San Antonio, San Antonio. Texas, U.S.A.
  • Departments of Biochemistry and Cellular and Structural Biology, University of Texas Health Science Center at San Antonio, San Antonio, Texas, U.S.A.
Venue
Journal of Bone and Mineral Research
Publisher
Oxford University Press (OUP)
Published
1997-12-01
Volume
12
Issue
12
Pages
2014-2023
ISSN
  • 0884-0431
  • 1523-4681
DOI
10.1359/jbmr.1997.12.12.2014
Cited by
500
References
0
Language
en
Licence
academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model (vor)

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