Transgenic expression of COL1A1-chloramphenicol acetyltransferase fusion genes in bone: differential utilization of promoter elements in vivo and in cultured cells

1993 ◽  
Vol 13 (9) ◽  
pp. 5168-5174
Author(s):  
P H Krebsbach ◽  
J R Harrison ◽  
A C Lichtler ◽  
C O Woody ◽  
D W Rowe ◽  
...  
Keyword(s):  
Cell Cultures ◽  
Collagen Synthesis ◽  
Cultured Cells ◽  
Bone Cells ◽  
Chloramphenicol Acetyltransferase ◽  
Bone Cell ◽  
Mrna Levels ◽  
Fusion Genes ◽  
Primary Bone ◽  
Bone Cell Cultures

To directly compare the patterns of collagen promoter expression in cells and tissues, the activity of COL1A1 fusion genes in calvariae of neonatal transgenic mice and in primary bone cell cultures derived by sequential digestion of transgenic calvariae was measured. ColCAT3.6 contains 3.6 kb (positions -3521 to +115) of the rat COL1A1 gene ligated to the chloramphenicol acetyltransferase (CAT) reporter gene. ColCAT2.3 and ColCAT1.7 are 5' deletion mutants which contain 2,296 and 1,672 bp, respectively, of COL1A1 DNA upstream from the transcription start site. ColCAT3.6 activity was 4- to 6-fold lower in primary bone cell cultures than in intact calvariae, while ColCAT2.3 activity was at least 100-fold lower in primary bone cells than in calvariae. These changes were accompanied by a threefold decrease in collagen synthesis and COL1A1 mRNA levels in primary bone cells compared with collagen synthesis and COL1A1 mRNA levels in freshly isolated calvariae. ColCAT3.6 and ColCAT2.3 activity was maintained in calvariae cultured in the presence or absence of serum for 4 to 7 days. Thus, when bone cells are removed from their normal microenvironment, there is parallel downregulation of collagen synthesis, collagen mRNA levels, and ColCAT3.6 activity, with a much greater decrease in ColCAT2.3. These data suggest that a 624-bp region of the COL1A1 promoter between positions -2296 and -1672 is active in intact and cultured bone but inactive in cultured cells derived from the bone. We suggest that the downregulation of COL1A1 activity in primary bone cells may be due to the loss of cell shape or to alterations in cell-cell and/or cell-matrix interactions that normally occur in intact bone.

scholarly journals Transgenic expression of COL1A1-chloramphenicol acetyltransferase fusion genes in bone: differential utilization of promoter elements in vivo and in cultured cells.

1993 ◽  
Vol 13 (9) ◽  
pp. 5168-5174 ◽  
Author(s):  
P H Krebsbach ◽  
J R Harrison ◽  
A C Lichtler ◽  
C O Woody ◽  
D W Rowe ◽  
...  
Keyword(s):  
Cell Cultures ◽  
Collagen Synthesis ◽  
Cultured Cells ◽  
Bone Cells ◽  
Chloramphenicol Acetyltransferase ◽  
Bone Cell ◽  
Mrna Levels ◽  
Fusion Genes ◽  
Primary Bone ◽  
Bone Cell Cultures

To directly compare the patterns of collagen promoter expression in cells and tissues, the activity of COL1A1 fusion genes in calvariae of neonatal transgenic mice and in primary bone cell cultures derived by sequential digestion of transgenic calvariae was measured. ColCAT3.6 contains 3.6 kb (positions -3521 to +115) of the rat COL1A1 gene ligated to the chloramphenicol acetyltransferase (CAT) reporter gene. ColCAT2.3 and ColCAT1.7 are 5' deletion mutants which contain 2,296 and 1,672 bp, respectively, of COL1A1 DNA upstream from the transcription start site. ColCAT3.6 activity was 4- to 6-fold lower in primary bone cell cultures than in intact calvariae, while ColCAT2.3 activity was at least 100-fold lower in primary bone cells than in calvariae. These changes were accompanied by a threefold decrease in collagen synthesis and COL1A1 mRNA levels in primary bone cells compared with collagen synthesis and COL1A1 mRNA levels in freshly isolated calvariae. ColCAT3.6 and ColCAT2.3 activity was maintained in calvariae cultured in the presence or absence of serum for 4 to 7 days. Thus, when bone cells are removed from their normal microenvironment, there is parallel downregulation of collagen synthesis, collagen mRNA levels, and ColCAT3.6 activity, with a much greater decrease in ColCAT2.3. These data suggest that a 624-bp region of the COL1A1 promoter between positions -2296 and -1672 is active in intact and cultured bone but inactive in cultured cells derived from the bone. We suggest that the downregulation of COL1A1 activity in primary bone cells may be due to the loss of cell shape or to alterations in cell-cell and/or cell-matrix interactions that normally occur in intact bone.

Influence of experimental conditions on osteoblast activity in human primary bone cell cultures

1990 ◽  
Vol 5 (4) ◽  
pp. 337-343 ◽  
Author(s):  
Pascale M. Chavassieux ◽  
Chantal Chenu ◽  
Alexandre Valentin-Opran ◽  
Blandine Merle ◽  
Pierre D. Delmas ◽  
...  
Keyword(s):  
Cell Cultures ◽  
Bone Cell ◽  
Experimental Conditions ◽  
Osteoblast Activity ◽  
Primary Bone ◽  
Bone Cell Cultures

Response of normal and osteoporotic human bone cells to mechanical stress in vitro

1998 ◽  
Vol 274 (6) ◽  
pp. E1113-E1120 ◽  
Author(s):  
Jozien G. H. Sterck ◽  
Jenneke Klein-Nulend ◽  
Paul Lips ◽  
Elisabeth H. Burger
Keyword(s):  
Nitric Oxide ◽  
Mechanical Stress ◽  
Cell Cultures ◽  
Bone Cells ◽  
Bone Cell ◽  
Human Bone ◽  
Animal Bone ◽  
Bone Cell Cultures

Bone adapts to mechanical stress, and bone cell cultures from animal origin have been shown to be highly sensitive to mechanical stress in vitro. In this study, we tested whether bone cell cultures from human bone biopsies respond to stress in a similar manner as animal bone cells and whether bone cells from osteoporotic patients respond similarly to nonosteoporotic donors. Bone cell cultures were obtained as outgrowth from collagenase-stripped trabecular bone fragments from 17 nonosteoporotic donors between 7 and 77 yr of age and from 6 osteoporotic donors between 42 and 72 yr of age. After passage, the cells were mechanically stressed by treatment with pulsating fluid flow (PFF; 0.7 ± 0.03 Pa at 5 Hz for 1 h) to mimic the stress-driven flow of interstitial fluid through the bone canaliculi, which is likely the stimulus for mechanosensation in bone in vivo. Similar to earlier studies in rodent and chicken bone cells, the bone cells from nonosteoporotic donors responded to PFF with enhanced release of prostaglandin E2(PGE2) and nitric oxide as well as a reduced release of transforming growth factor-β (TGF-β). The upregulation of PGE2 but not the other responses continued for 24 h after 1 h of PFF treatment. The bone cells from osteoporotic donors responded in a similar manner as the nonosteoporotic donors except for the long-term PGE2 release. The PFF-mediated upregulation of PGE2 release during 24 h of postincubation after 1 h of PFF was significantly reduced in osteoporotic patients compared with six age-matched controls as well as with the whole nonosteoporotic group. These results indicate that enhanced release of PGE2 and nitric oxide, as well as reduced release of TGF-β, is a characteristic response of human bone cells to fluid shear stress, similar to animal bone cells. The results also suggest that bone cells from osteoporotic patients may be impaired in their long-term response to mechanical stress.

scholarly journals Effects of Low Intensity Pulsed Ultrasound And Low Level Heat on Bone Cells

2021 ◽  
Author(s):  
Judith Weidman
Keyword(s):  
Cell Cultures ◽  
Bone Cells ◽  
Bone Cell ◽  
Pulsed Ultrasound ◽  
Alizarin Red ◽  
Treatment Groups ◽  
Low Level ◽  
Low Intensity Pulsed Ultrasound ◽  
Low Intensity ◽  
Bone Cell Cultures

Both Low Intensity Pulsed UltraSound (LIPUS) and low level heat have separately been shown to improve mineralization in bone cell cultures (Unsworth et al 2007, Leon et al 1993). This study examines the effect of concurrent LIPUS and low level heat on MC3T3-E1 bone cell cultures. The treatment groups were: LIPUS, heat, LIPUS + heat, and control. The LIPUS intensity was ISATA=10 mW/cm2 at f=1.5 MHz, and heat was applied at 40ºC for 40 minutes each day over 15 days. The LIPUS + heat group received the treatments concurrently. The groups were compared using Alizarin Red staining to measure the degree of cell mineralization. All treatment groups showed statistically significantly improved mineralization over the control; however, there was no statistical difference between the LIPUS and the LIPUS + heat groups. Early results suggest that concurrent heat and LIPUS on MC3T3-E1 bone cells has no additive effect on mineralization.

PPAR agonists stimulate adipogenesis at the expense of osteoblast differentiation in primary bone cell cultures

Bone ◽  
2011 ◽  
Vol 48 ◽  
pp. S112-S113
Author(s):  
J.J. Patel⁎ ◽  
O.R. Butters ◽  
S.E.B. Taylor ◽  
T.R. Arnett
Keyword(s):  
Cell Cultures ◽  
Osteoblast Differentiation ◽  
Bone Cell ◽  
Ppar Agonists ◽  
Primary Bone ◽  
Bone Cell Cultures

Differential solubilization of osteoblastic alkaline phosphatase from human primary bone cell cultures

1996 ◽  
Vol 28 (4) ◽  
pp. 421-430 ◽  
Author(s):  
Jacqueline Radisson ◽  
Muriel Angrand ◽  
Pascale Chavassjeux ◽  
Bernard Roux ◽  
Gérard Azzar
Keyword(s):  
Alkaline Phosphatase ◽  
Cell Cultures ◽  
Bone Cell ◽  
Primary Bone ◽  
Bone Cell Cultures

scholarly journals Effects of Low Intensity Pulsed Ultrasound And Low Level Heat on Bone Cells

2021 ◽  
Author(s):  
Judith Weidman
Keyword(s):  
Cell Cultures ◽  
Bone Cells ◽  
Bone Cell ◽  
Pulsed Ultrasound ◽  
Alizarin Red ◽  
Treatment Groups ◽  
Low Level ◽  
Low Intensity Pulsed Ultrasound ◽  
Low Intensity ◽  
Bone Cell Cultures

Both Low Intensity Pulsed UltraSound (LIPUS) and low level heat have separately been shown to improve mineralization in bone cell cultures (Unsworth et al 2007, Leon et al 1993). This study examines the effect of concurrent LIPUS and low level heat on MC3T3-E1 bone cell cultures. The treatment groups were: LIPUS, heat, LIPUS + heat, and control. The LIPUS intensity was ISATA=10 mW/cm2 at f=1.5 MHz, and heat was applied at 40ºC for 40 minutes each day over 15 days. The LIPUS + heat group received the treatments concurrently. The groups were compared using Alizarin Red staining to measure the degree of cell mineralization. All treatment groups showed statistically significantly improved mineralization over the control; however, there was no statistical difference between the LIPUS and the LIPUS + heat groups. Early results suggest that concurrent heat and LIPUS on MC3T3-E1 bone cells has no additive effect on mineralization.

Effect of Angiotensin II on Inflammation Pathways in Human Primary Bone Cell Cultures in Otosclerosis

2012 ◽  
Vol 17 (3) ◽  
pp. 169-178 ◽  
Author(s):  
Milan Rudić ◽  
Christine Nguyen ◽  
Yann Nguyen ◽  
Lidija Milković ◽  
Neven Žarković ◽  
...  
Keyword(s):  
Angiotensin Ii ◽  
Cell Cultures ◽  
Bone Cell ◽  
Primary Bone ◽  
Bone Cell Cultures
Sign in / Sign up

Export Citation Format

Share Document