Mineralization induced by beta-glycerophosphate in cultures leads to a marked increase in collagenase synthesis by mouse osteogenic MC3T3-E1 cells under subsequent stimulation with heparin.
The clonally derived mouse osteoblast-like cell line MC3T3-E1 was shown to produce latent collagenase (approximately 0.2 units/ml) under stimulation with either heparin or parathyroid hormone in confluent cultures. However, it was found that MC3T3 E1 cultures which were first induced to undergo mineralization by the addition of beta-glycerophosphate and were subsequently stimulated with heparin showed an approximately ten-fold increase in collagenase synthesis. MC3T3-E1 cell collagenase from a small sample of serum-free culture medium was purified 49-fold to a specific activity of 200 units/mg protein with a yield of 14% by heparin-sepharose affinity chromatography and ion-exchange high performance liquid chromatography. This new mineralization-primed cell culture system may be a valuable model for the study of osteoblast collagenase.
Sakamoto
S
S
Laboratory for the Study of Connective Tissue Metabolism, Harvard School of Dental Medicine, Boston, Massachusetts.
Sakamoto
M
M
Goldberg
L
L
Colarusso
L
L
Gotoh
Y
Y
D000469:Alkaline Phosphatase / Q000096:biosynthesis; D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005994:Glycerophosphates / Q000494:pharmacology*; D006493:Heparin / Q000494:pharmacology*; D051379:Mice; D003012:Microbial Collagenase / Q000096:biosynthesis*; D008903:Minerals / Q000378:metabolism*; D010006:Osteoblasts / Q000187:drug effects / Q000201:enzymology*; D010281:Parathyroid Hormone / Q000494:pharmacology
D005994:Glycerophosphates; D008903:Minerals; D010281:Parathyroid Hormone; D006493:Heparin; D000469:Alkaline Phosphatase; D003012:Microbial Collagenase; C031463:beta-glycerophosphoric acid