Abstract
Propagation in vitro of rat tibial osteoblasts (ROB) is accompanied by increased expression of the early osteogenic marker alkaline phosphatase (AP) and maturation of the osteogenic phenotype. In order to establish the pattern of the integrin expressed in ROB during progression to the mature osteoblastic phenotype, we have used biosynthetic, immunoblotting and immunohistochemical assays. We immunoprecipitated from osteoblasts, expanded for 1.5- and 7.5-doubling, α5β1, αvβ3, α3β1, α6β1 and α1β1 integrin heterodimers; furthermore β5, α2 and α4 chains were detected by immunoblots and indirect immunofluorescence, αv, α1, α6 subunits in most cells, and β3 and β1 subunits in a minority, were found to be associated with adhesion plaques in osteoblasts of 1.5-, 4.5- and 7.5-doubling grown in the presence of FCS, while all other subunits stained diffusely all the cells. Adhesion to fibronectin (FN), laminin (LN), collagen type I (COL I) and III (COL III) by ROB at different doubling (1.5-11) was dependent on substratum concentration, and after 2.5 h at 55 nM 60% of the cells adhered to all substrata. Arg-Gly-Asp-Ser (RGDS) containing peptides inhibited adhesion of cells differentially, according to substratum; no dependence on extent of progation in vitro was observed. In conclusion, ROB cultured in vitro for 1.5- to 11-doubling had an unchanged pattern of expression of integrin subunits, heterodimer association and cellular distribution. Adhesion specificity and affinity were also unchanged. These results suggest that the phenotypic maturation, detected as an increase in AP expression, is not accompanied by major changes in the potential for cell-matrix interactions, and does not correspond to changes in the type of integrin subunits expressed by osteoblasts.
| Original language | English |
|---|---|
| Pages (from-to) | 7-16 |
| Number of pages | 10 |
| Journal | Cell Biology International |
| Volume | 21 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - Jan 1997 |
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Keywords
- Adhesion
- In vitro propagation
- Integrins
- Osteoblasts
ASJC Scopus subject areas
- Cell Biology
Cite this
Osteoblastic cells from rat long bone II : Adhesion to substrata and integrin expression in primary and propagated cultures. / Castoldi, Mirco; Pistone, Mario; Caruso, Cristina; Puddu, Alessandra; Filanti, Cristina; Piccini, Daniele; Tacchetti, Carlo; Manduca, Paola.
In: Cell Biology International, Vol. 21, No. 1, 01.1997, p. 7-16.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Osteoblastic cells from rat long bone II
T2 - Adhesion to substrata and integrin expression in primary and propagated cultures
AU - Castoldi, Mirco
AU - Pistone, Mario
AU - Caruso, Cristina
AU - Puddu, Alessandra
AU - Filanti, Cristina
AU - Piccini, Daniele
AU - Tacchetti, Carlo
AU - Manduca, Paola
PY - 1997/1
Y1 - 1997/1
N2 - Propagation in vitro of rat tibial osteoblasts (ROB) is accompanied by increased expression of the early osteogenic marker alkaline phosphatase (AP) and maturation of the osteogenic phenotype. In order to establish the pattern of the integrin expressed in ROB during progression to the mature osteoblastic phenotype, we have used biosynthetic, immunoblotting and immunohistochemical assays. We immunoprecipitated from osteoblasts, expanded for 1.5- and 7.5-doubling, α5β1, αvβ3, α3β1, α6β1 and α1β1 integrin heterodimers; furthermore β5, α2 and α4 chains were detected by immunoblots and indirect immunofluorescence, αv, α1, α6 subunits in most cells, and β3 and β1 subunits in a minority, were found to be associated with adhesion plaques in osteoblasts of 1.5-, 4.5- and 7.5-doubling grown in the presence of FCS, while all other subunits stained diffusely all the cells. Adhesion to fibronectin (FN), laminin (LN), collagen type I (COL I) and III (COL III) by ROB at different doubling (1.5-11) was dependent on substratum concentration, and after 2.5 h at 55 nM 60% of the cells adhered to all substrata. Arg-Gly-Asp-Ser (RGDS) containing peptides inhibited adhesion of cells differentially, according to substratum; no dependence on extent of progation in vitro was observed. In conclusion, ROB cultured in vitro for 1.5- to 11-doubling had an unchanged pattern of expression of integrin subunits, heterodimer association and cellular distribution. Adhesion specificity and affinity were also unchanged. These results suggest that the phenotypic maturation, detected as an increase in AP expression, is not accompanied by major changes in the potential for cell-matrix interactions, and does not correspond to changes in the type of integrin subunits expressed by osteoblasts.
AB - Propagation in vitro of rat tibial osteoblasts (ROB) is accompanied by increased expression of the early osteogenic marker alkaline phosphatase (AP) and maturation of the osteogenic phenotype. In order to establish the pattern of the integrin expressed in ROB during progression to the mature osteoblastic phenotype, we have used biosynthetic, immunoblotting and immunohistochemical assays. We immunoprecipitated from osteoblasts, expanded for 1.5- and 7.5-doubling, α5β1, αvβ3, α3β1, α6β1 and α1β1 integrin heterodimers; furthermore β5, α2 and α4 chains were detected by immunoblots and indirect immunofluorescence, αv, α1, α6 subunits in most cells, and β3 and β1 subunits in a minority, were found to be associated with adhesion plaques in osteoblasts of 1.5-, 4.5- and 7.5-doubling grown in the presence of FCS, while all other subunits stained diffusely all the cells. Adhesion to fibronectin (FN), laminin (LN), collagen type I (COL I) and III (COL III) by ROB at different doubling (1.5-11) was dependent on substratum concentration, and after 2.5 h at 55 nM 60% of the cells adhered to all substrata. Arg-Gly-Asp-Ser (RGDS) containing peptides inhibited adhesion of cells differentially, according to substratum; no dependence on extent of progation in vitro was observed. In conclusion, ROB cultured in vitro for 1.5- to 11-doubling had an unchanged pattern of expression of integrin subunits, heterodimer association and cellular distribution. Adhesion specificity and affinity were also unchanged. These results suggest that the phenotypic maturation, detected as an increase in AP expression, is not accompanied by major changes in the potential for cell-matrix interactions, and does not correspond to changes in the type of integrin subunits expressed by osteoblasts.
KW - Adhesion
KW - In vitro propagation
KW - Integrins
KW - Osteoblasts
UR - http://www.scopus.com/inward/record.url?scp=0031048739&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0031048739&partnerID=8YFLogxK
U2 - 10.1006/cbir.1996.0110
DO - 10.1006/cbir.1996.0110
M3 - Article
C2 - 9046103
AN - SCOPUS:0031048739
VL - 21
SP - 7
EP - 16
JO - Cell Biology International
JF - Cell Biology International
SN - 1065-6995
IS - 1
ER -