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  • Tissue culture  (2)
  • Springer  (2)
  • Public Library of Science
  • 1
    Electronic Resource
    Electronic Resource
    Effect of multiplication-stimulating activity on DNA and protein synthesis in cultured fetal rat calvaria (1979)
    Springer
    Calcified tissue international 29 (1979), S. 33-39 
    Details
    ISSN: 1432-0827
    Keywords: Multiplication-stimulating activity ; Somatomedin ; Tissue culture ; Bone formation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Multiplication-stimulating Activity (MSA), a peptide closely related to somatomedin, was examined for its effects on bone formation and sulfation activity in vitro. To study the effect of MSA on bone formation, we examined its effects on the incorporation of3H-thymidine into DNA,3H-uridine into RNA, and3H-proline into collagenase-digestible (CDP) and noncollagen protein (NCP), as well as DNA content in 21-day fetal rat calvaria cultured for 24 h periods. MSA caused a dose-dependent stimulation of the incorporation of3H-thymidine into DNA at concentrations of 1 to 3µg/ml, increased the total DNA content, and had no effect on3H-uridine incorporation. The effect on3H-thymidine incorporation appeared and was maximal at 12 h and was sustained for 24 h. After 24 h of treatment, MSA, 1 to 3µg/ml, caused a 25 to 60% stimulation on the incorporation of3H-proline into CDP and NCP. The effect was not specific for CDP, and there was no increase in the percent collagen synthesized. In contrast, insulin, 10 nM, increased the labeling of CDP by 95% but had a small effect on NCP and did not affect DNA synthesis. Insulin did not diminish the MSA effect on CDP and NCP labeling but slightly decreased its effect on thymidine incorporation. Cortisol inhibited DNA synthesis, but MSA was effective in its presence and cortisol did not affect the stimulatory effect of MSA on CDP or NCP. Histological sections showed a sixfold increase in the mitotic index after colcemid arrest in MSA-treated bones. MSA, 0.1 to 5µg/ml, also stimulated the incorporation of35S into pig costal cartilage in culture. Our studies indicate that MSA stimulates DNA, collagen, and noncollagen protein synthesis in bone cultures and stimulates sulfate incorporation into cartilage.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02408053
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  • 2
    Electronic Resource
    Electronic Resource
    Effect of sex steroids on bone collagen synthesis in vitro (1978)
    Springer
    Calcified tissue international 25 (1978), S. 105-110 
    Details
    ISSN: 1432-0827
    Keywords: Progesterone ; 17β estradiol ; Testosterone ; Tissue culture ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Although sex steroids are known to affect skeletal metabolism, their effects on bone collagen synthesis have not been studied. We have examined the direct effects of progesterone, 17β estradiol, testosterone, 5α dihydrotestosterone and dehydroepiandrosterone on bone collagen and noncollagen protein synthesis in cultures of calvaria obtained from 21-day fetal rats. Bones were incubated for 24 to 168 h and3H-proline was added for the last 2 h of culture. Incorporation of the label into collagenase-digestible protein (CDP)1 and noncollagen protein (NCP) was determined using repurified bacterial collagenase. Progesterone caused a dose dependent inhibition of the labeling of CDP at concentrations of 3×10−7 M to 10−5 M after 96 h of culture. A smaller inhibitory effect was observed on NCP. The inhibitory effect was slow in onset and persisted when bones were incubated for 48 h with progesterone and then transferred to control medium for 48 h. Progesterone also inhibited the incorporation of3H-thymidine and3H-uridine into fetal rat calvaria. After 24 h of culture, 17β estradiol and testosterone had no effects on the labeling of CDP and NCP. After 96 h, 17β estradiol had a small and inconsistent stimulatory effect on the labeling of CDP but testosterone had no effect. Neither hormone altered the inhibitory effects of parathyroid hormone, cortisol and progesterone. Dihydrotestosterone and dehydroepiandrosterone had no effects after 24 h and 96 h of culture. 17β estradiol, testosterone and dihydrotestosterone did not affect the incorporation of3H-uridine or3H-thymidine into fetal rat calvaria. Our studies indicate that progesterone is an inhibitor of bone collagen synthesis and estrogens and androgens are not major regulators of bone formation in vitro.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02010758
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