Gene and protein expression during differentiation and matrix mineralization in a chondrocyte cell culture system

Calcified Tissue International
N KergosienN Forest

Abstract

Endochondral bone formation occurs through a series of developmentally regulated cellular stages, from initial formation of cartilage tissue to calcified cartilage, resorption, and replacement by bone tissue. Nasal cartilage cells isolated by enzymatic digestion from rat fetuses were seeded at a final density of 10(5) cell/cm2 and cultured in Dulbecco's modified Eagle medium (DMEM) supplemented with 10% fetal calf serum in the presence of ascorbic acid and beta-glycerophosphate. First, cells lost their phenotype but in this condition they rapidly reexpressed the chondrocyte phenotype and were able to form calcified cartilaginous nodules with the morphological appearance of cartilage mineralization that occurs in vivo during endochondral ossification. In this mineralizing chondrocyte culture system, we investigated, between day 3 and day 15, the pattern expression of types II and X collagen, proteoglycan core protein, characteristic markers of chondrocyte differentiation, as well as alkaline phosphatase and osteocalcin associated with the mineralization process. Analysis of labeled collagen and immunoblotting revealed type I collagen synthesis associated with the loss of chondrocyte phenotype at the beginning of the culture. How...Continue Reading

Citations

Sep 10, 2010·Tissue Engineering. Part B, Reviews·Karym El SayedGundula Schulze-Tanzil
Nov 26, 1999·Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research·Y Sun, R Kandel
Mar 9, 2006·Calcified Tissue International·L F A HuitemaC H A van de Lest
Mar 26, 2004·Archives of Oral Biology·G O Ramirez-YañezW G Young
Feb 18, 2015·Journal of Cellular and Molecular Medicine·Benjamin E PippengerIvan Martin
Aug 21, 2008·Nature Protocols·Marjolaine GossetClaire Jacques
Mar 29, 2002·Journal of Orthopaedic Research : Official Publication of the Orthopaedic Research Society·Roy K AaronDeborah M Ciombor

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