Subcloning of three osteoblastic cell lines with distinct differentiation phenotypes from the mouse osteoblastic cell line KS-4

Yamashita, Takefumi; Ishii, Hiromi; Shimoda, Kei; Sampath, T. Kuber; Katagiri, Takenobu; Wada, Michihito; Osawa, Tatsushi; Suda, Toshio

doi:10.1016/s8756-3282(96)00255-4

Cited by 39 publications

(27 citation statements)

References 38 publications

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“…This differentiation profile is similar to that of other species (Robey & Termine 1985, Marie et al 1989, Owen et al 1990. The increase in COL1 mRNA expression observed during the later stages of osteoblast differentiation is consistent with a number of other studies in differentiated mouse, but not rat, osteoblasts (Owen et al 1991, Yamashita et al 1996, Lian et al 1997, Qu et al 1998.…”

Section: Discussionsupporting

confidence: 90%

Changing RANKL/OPG mRNA expression in differentiating murine primary osteoblasts

Thomas

Baker

Eisman³

et al. 2001

Journal of Endocrinology

164

114

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Osteoblast-osteoclast coordination is critical in the maintenance of skeletal integrity. The modulation of osteoclastogenesis by immature cells of the osteoblastic lineage is mediated through receptor activator of NF B (RANK), its ligand RANKL, and osteoprotegerin (OPG), a natural decoy receptor for RANKL. Here, the expression of OPG and RANKL in primary mouse osteoblastic cultures was investigated to determine whether the osteoclastogenic stimulus depended on the stage of osteoblastic differentiation and the presence of the calciotrophic hormone 1,25OPG mRNA expression was increased in osteoblastic cultures after the onset of mineralisation relative to less mature cultures, but did not alter in response to 1,25-(OH) 2 D 3 treatment. In contrast, basal RANKL mRNA expression did not change during differentiation but was significantly enhanced by 1,25-(OH) 2 D 3 treatment at all times. The stimulatory effects of 1,25-(OH) 2 D 3 on RANKL were lessened in more mature cultures, however. The RANKL/OPG ratio, an index of osteoclastogenic stimulus, was therefore increased by 1,25-(OH) 2 D 3 treatment at all stages of osteoblastic differentiation, but to a lesser degree in cultures after the onset of mineralisation. Thus the 1,25-(OH) 2 D 3 -driven increase in osteoclastogenic potential of immature osteoblasts appears to be mediated by increased RANKL mRNA expression, with mature osteoblasts having relatively decreased osteoclastogenic activity due to increased OPG mRNA expression. These findings suggest a possible mechanism for the recently proposed negative regulatory role of mature osteoblasts on osteoclastogenesis and indicate that the relative proportions of immature and mature osteoblasts in the local microenvironment may control the degree of resorption at each specific bone site.

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Section: Discussionsupporting

confidence: 90%

Changing RANKL/OPG mRNA expression in differentiating murine primary osteoblasts

Thomas

Baker

Eisman³

et al. 2001

Journal of Endocrinology

164

114

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“…(38) KS483 is a fetal mouse calvaria-derived clonal cell line committed to the osteoblast lineage. (28,29) The finding that these cells could differentiate into both osteoblasts and adipocytes shows that this cell line still has progenitor characteristics. In both KS483 and mouse mesenchymal bone marrow cultures, cells first proliferate and then differentiate into either osteoblasts that form calcified bone nodules or adipocytes.…”

Section: Discussionmentioning

confidence: 99%

“…For this, we used KS483 cells, which is a clonal cell line from fetal mouse calvaria. (28,29) These cells are able to form both calcified nodules and adipocytes when they are cultured in charcoal-stripped fetal bovine serum (FBS). (30) In addition, we studied the effects of estrogen on osteogenesis and adipogenesis in mouse bone marrow cells.…”

mentioning

confidence: 99%

Exposure of KS483 Cells to Estrogen Enhances Osteogenesis and Inhibits Adipogenesis

Dang

Bezooijen

Karperien

et al. 2002

Journal of Bone and Mineral Research

166

118

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Osteoblasts and adipocytes arise from a common progenitor cell in bone marrow. Whether estrogen directly regulates the progenitor cells differentiating into osteoblasts or adipocytes remains unknown. Using a mouse clonal cell line KS483 cultured in charcoal-stripped fetal bovine serum (FBS), we showed that 17␤-estradiol (E 2 ) stimulates the differentiation of progenitor cells into osteoblasts and concurrently inhibits adipocyte formation in an estrogen receptor (ER)-dependent way. E 2 increased alkaline phosphate (ALP) activity and nodule formation and stimulated messenger RNA (mRNA) expression of core-binding factor ␣-1 (Cbfa1), parathyroid hormone/parathyroid hormone-related protein receptors (PTH/PTHrP-Rs), and osteocalcin. In contrast, E 2 decreased adipocyte numbers and down-regulated mRNA expression of peroxisome proliferatoractivated receptor-␥ (PPAR␥)2, adipocyte protein 2 (aP2), and lipoprotein lipase (LPL). Furthermore, the reciprocal control of osteoblast and adipocyte differentiation by E 2 was observed also in the presence of the adipogenic mixture of isobutylmethylxanthine, dexamethasone, and insulin. Immunohistochemical staining showed that ER␣ and ER␤ were present in osteoblasts and adipocytes. A new mouse splice variant ER␤2 was identified, which differed in two amino acid residues from the rat isoform.

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“…TRIzol reagent (Invitrogen) was used to extract RNA from 6-dpf embryos, and mouse RNA was extracted from cultured KS483 cells (34). For details, please consult SI Materials and Methods.…”

Section: Methodsmentioning

confidence: 99%

Entpd5 is essential for skeletal mineralization and regulates phosphate homeostasis in zebrafish

Huitema

Apschner

Logister

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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Bone mineralization is an essential step during the embryonic development of vertebrates, and bone serves vital functions in human physiology. To systematically identify unique gene functions essential for osteogenesis, we performed a forward genetic screen in zebrafish and isolated a mutant, no bone (nob), that does not form any mineralized bone. Positional cloning of nob identified the causative gene to encode ectonucleoside triphosphate/ diphosphohydrolase 5 (entpd5); analysis of its expression pattern demonstrates that entpd5 is specifically expressed in osteoblasts. An additional mutant, dragonfish (dgf), exhibits ectopic mineralization in the craniofacial and axial skeleton and encodes a loss-offunction allele of ectonucleotide pyrophosphatase phosphodiesterase 1 (enpp1). Intriguingly, generation of double-mutant nob/ dgf embryos restored skeletal mineralization in nob mutants, indicating that mechanistically, Entpd5 and Enpp1 act as reciprocal regulators of phosphate/pyrophosphate homeostasis in vivo. Consistent with this, entpd5 mutant embryos can be rescued by high levels of inorganic phosphate, and phosphate-regulating factors, such as fgf23 and npt2a, are significantly affected in entpd5 mutant embryos. Our study demonstrates that Entpd5 represents a previously unappreciated essential player in phosphate homeostasis and skeletal mineralization.

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Subcloning of three osteoblastic cell lines with distinct differentiation phenotypes from the mouse osteoblastic cell line KS-4

Cited by 39 publications

References 38 publications

Changing RANKL/OPG mRNA expression in differentiating murine primary osteoblasts

Changing RANKL/OPG mRNA expression in differentiating murine primary osteoblasts

Exposure of KS483 Cells to Estrogen Enhances Osteogenesis and Inhibits Adipogenesis

Entpd5 is essential for skeletal mineralization and regulates phosphate homeostasis in zebrafish

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