Establishment of human osteoblastic cells derived from periosteum in culture |
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Authors: | Yasuko Koshihara Mieko Kawamura Sachiko Endo Chiharu Tsutsumi Hiroaki Kodama Hiromi Oda Shohzo Higaki |
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Institution: | (1) Department of Pharmacology, Tokyo Metropolitan Institute of Gerontology, Itabashi-ku, 173 Tokyo, Japan;(2) Department of Biophysics and Biochemistry, Faculty of Science, University of Tokyo, Bunkyo-ku, 173 Tokyo, Japan;(3) Department of Orthopaedic Surgery Faculty of Medicine, University of Tokyo, Bunkyo-ku, 173 Tokyo, Japan |
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Abstract: | Summary We isolated osteoblastic cells derived from human periosteum and established them in culture. Their growth depended on the
presence of ascorbic acid, and the doubling time was 40 to 60 h. The requirement for ascorbic acid was used to high production
of collagen. These cells produced mainly type I collagen and only small amounts of type III collagen determined by reducing
sodium dodecyl sulfate SDS-polyacrylamide gel electrophoresis. The total collagen yield was about 10 mg from 2×107 cells. The cells could be continuously cultured in α-minimum essential medium supplemented with 10% fetal bovine serum for
18 to 40 population doubling levels, depending on the age of the donated periosteum. These cells have the ability to calcify
when incubated with 2 mM α-glycerophosphate-Na2. Calcification as viewed by the naked eye appeared from Day 15 after treatment. Treatment with the active formed vitamin
D3, 1, 25 dihydroxyvitamin D3 enhanced calcification significantly and stimulated osteocalcin production. By electron microscopy, cells with many projections
on their surfaces showed well-developed rough endoplasmic reticulum and actinlike fibers, and large numbers of lysosomes,
mitochondria, and secretion granules. Many matrix vesicles, in which minerals were initially localized, and well-banded collagen
fibrils were seen in the intercellular spaces. These observations demonstrate typical osteoblastic morphology. The above results
indicate that cultured cells from human periostem are osteoblastic cells that have the capacity to differentiate into osteocytes
and to deposit calcified minerals in response to 1,25 dihydroxyvitamin D3. |
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Keywords: | osteoblastic cells calcification human osteocalcin alkaline phosphatase vitamin D3 |
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