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Am J Physiol Cell Physiol 279: C1220-C1229, 2000;
0363-6143/00 $5.00
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Vol. 279, Issue 4, C1220-C1229, October 2000

Bioenergetics and mitochondrial transmembrane potential during differentiation of cultured osteoblasts

Svetlana V. Komarova1, Fasoil I. Ataullakhanov2, and Ruth K. Globus1

1 Life Sciences Division, NASA Ames Research Center, Moffett Field, California 94035-1000; and 2 National Research Center for Hematology, Moscow, 125167 Russia

To evaluate the relationship between osteoblast differentiation and bioenergetics, cultured primary osteoblasts from fetal rat calvaria were grown in medium supplemented with ascorbate to induce differentiation. Before ascorbate treatment, the rate of glucose consumption was 320 nmol · h-1 · 106 cells-1, respiration was 40 nmol · h-1 · 106 cells-1, and the ratio of lactate production to glucose consumption was ~2, indicating that glycolysis was the main energy source for immature osteoblasts. Ascorbate treatment for 14 days led to a fourfold increase in respiration, a threefold increase in ATP production, and a fivefold increase in ATP content compared with that shown in immature cells. Confocal imaging of mitochondria stained with a transmembrane potential-sensitive vital dye showed that mature cells possessed abundant amounts of high-transmembrane-potential mitochondria, which were concentrated near the culture medium-facing surface. Acute treatment of mature osteoblasts with metabolic inhibitors showed that the rate of glycolysis rose to maintain the cellular energy supply constant. Thus progressive differentiation coincided with changes in cellular metabolism and mitochondrial activity, which are likely to play key roles in osteoblast function.

ATP; respiration; glycolysis; bone


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