The influence of culture conditions on extracellular matrix proteins synthesized by osteoblasts derived from rabbit bone marrow.
The influence of culture conditions on the extracellular matrix (ECM) protein expressions of rabbit bone marrow stromal cells has been studied. The focus was on the effects of two kinds of sera, fetal calf serum (FCS) and Ultroser, on cells treated with dexamethasone. The induction of osteoblastic differentiation by dexamethasone addition is confirmed, particularly when cells are cultured in FCS. Bone marrow stromal cells produce alkaline phosphatase positive CFU-F and produce ECM with some mineralized nodules. Analysis by means of two-dimensional gel electrophoresis showed important changes in the composition of ECM proteins after dexamethasone treatment. Overexpression, underexpression, and new synthesized proteins were observed. The most significant modification was linked to the synthesis of four new proteins visible in the acidic area with a low molecular weight of around 17 kDa. These proteins did not correspond to those ECM proteins known to be induced by dexamethasone. Moreover, the effect of dexamethasone on osteoblastic differentiation induction appears very limited when cells are cultured in Ultroser compared to FCS. The protein pattern with Ultroser is different to that obtained with FCS. Cells cultured in Ultroser synthesized no new protein. The different behavior of cells according to the type of medium used is discussed in terms of the osteogenic factors present in the two different sera.
Institut de Recherche sur les Biomatériaux et les Biotechnologies (IR2B), Université du Littoral Côte d'Opale, Bassin Napoléon, Quai Robert Masset, BP 120 62327 Boulogne sur mer, France.
François
F
Cornet
Karine
K
Anselme
Thierry
T
Grard
Myriam
M
Rouahi
Benoit
B
Noël
Pierre
P
Hardouin
Joseph
J
Jeanfils
D000818:Animals; D001854:Bone Marrow Cells; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects; D003470:Culture Media / Q000494:pharmacology; D003907:Dexamethasone / Q000494:pharmacology; D015180:Electrophoresis, Gel, Two-Dimensional; D016326:Extracellular Matrix Proteins / Q000032:analysis / Q000187:drug effects / Q000378:metabolism*; D005260:Female; D010006:Osteoblasts / Q000378:metabolism*; D011506:Proteins / Q000032:analysis; D011817:Rabbits; D013234:Stem Cells / Q000166:cytology; D017154:Stromal Cells / Q000166:cytology
D003470:Culture Media; D016326:Extracellular Matrix Proteins; D011506:Proteins; D003907:Dexamethasone