Selective attachment of osteoprogenitors to laminin.
During endochondral ossification and bone remodeling, osteoprogenitors (OP) attach to the matrix and differentiate into osteoblasts. To identify matrix proteins binding specifically these precursors, fetal rat calvaria (RC) cells were plated for 5-20 min in serum-free medium, on wells coated with various proteins and saturated with bovine serum albumin (BSA) to block nonspecific binding sites. Adherent cells were either counted or grown to assess bone colony (nodule) formation. As each nodule originates from the clonal division of one OP, the ratio (nodules/100 cells attached) measures the proportion of OP among adherent cells. Of numerous purified matrix proteins tested, laminin-1 and tenascin inhibited cell attachment, whereas fibronectin, bone sialoprotein, and type I collagen increased cell attachment and others had no effect. Only laminin-1 and, to a lesser extent, tenascin, enriched the cell population in OP. Laminin-1 acted time- and dose-dependently. In experiments in which cell attachment to laminin-coated but unsaturated wells was ensured by plating for 24 h in 10% fetal calf serum, laminin-1 had no effect on cell attachment nor on OP differentiation. In contrast, repeated plating of RC cells on laminin-1-coated/saturated wells depleted the population in OP, confirming that OP selection was a cell-attachment effect. When RC cell populations isolated by successive collagenase extractions were compared, the highest rate of OP enrichment on laminin-1 was obtained with the earliest populations, which were the most responsive to dexamethasone, a marker of early OP stages. In conclusion, laminin-1 recruits in vitro, through a cell-attachment effect, OP present in early RC cell populations, of which laminins are abundant extracellular matrix components.
Roche
P
P
INSERM Unité 403, Hôpital E. Herriot, Lyon, France.
Goldberg
H A
HA
Delmas
P D
PD
Malaval
L
L
D000818:Animals; D002417:Cattle; D002448:Cell Adhesion / Q000502:physiology; D002478:Cells, Cultured; D016326:Extracellular Matrix Proteins / Q000032:analysis / Q000502:physiology; D007797:Laminin / Q000502:physiology*; D010006:Osteoblasts / Q000502:physiology*; D051381:Rats; D013234:Stem Cells / Q000502:physiology*
D016326:Extracellular Matrix Proteins; D007797:Laminin