PMID- 11204431
OWN - NLM
STAT- MEDLINE
DCOM- 20010419
LR  - 20131121
IS  - 0884-0431 (Print)
IS  - 0884-0431 (Linking)
VI  - 16
IP  - 2
DP  - 2001 Feb
TI  - A novel cell culture model of chondrocyte differentiation during mammalian 
      endochondral ossification.
PG  - 309-18
AB  - Endochondral ossification (EO) occurs in the growth plate where chondrocytes pass 
      through discrete stages of proliferation, maturation, hypertrophy, and 
      calcification. We have developed and characterized a novel bovine cell culture 
      model of EO that mirrors these events and will facilitate in vitro studies on 
      factors controlling chondrocyte differentiation. Chondrocytes derived from the 
      epiphyses of long bones of fetal calves were treated with 5-azacytidine (aza-C) 
      for 48 h. Cultures were maintained subsequently without aza-C and harvested at 
      selected time points for analyses of growth and differentiation status. A 
      chondrocytic phenotype associated with an extensive extracellular matrix rich in 
      proteoglycans and collagen types II and VI was observed in aza-C-treated and 
      -untreated cultures. aza-C-treated cultures were characterized by studying the 
      expression of several markers of chondrocyte differentiation. Parathyroid 
      hormone-related protein (PTHrP) and its receptor, both markers of maturation, 
      were expressed at days 5-9. Type X collagen, which is restricted to the stage of 
      hypertrophy, was expressed from day 11 onward. Hypertrophy was confirmed by a 
      14-fold increase in cell size by day 15 and an increased synthesis of alkaline 
      phosphatase during the hypertrophic period (days 14-28). The addition of PTHrP to 
      aza-C-treated cultures at day 14 led to the down-regulation of type X collagen by 
      6-fold, showing type X collagen expression is under the control of PTHrP as in 
      vivo. These findings show that aza-C can induce fetal bovine epiphyseal 
      chondrocytes to differentiate in culture in a manner consistent with that which 
      occurs during the EO process in vivo.
FAU - Cheung, J O
AU  - Cheung JO
AD  - Wellcome Trust Center for Cell-Matrix Research, School of Biological Sciences, 
      University of Manchester, UK.
FAU - Hillarby, M C
AU  - Hillarby MC
FAU - Ayad, S
AU  - Ayad S
FAU - Hoyland, J A
AU  - Hoyland JA
FAU - Jones, C J
AU  - Jones CJ
FAU - Denton, J
AU  - Denton J
FAU - Thomas, J T
AU  - Thomas JT
FAU - Wallis, G A
AU  - Wallis GA
FAU - Grant, M E
AU  - Grant ME
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - J Bone Miner Res
JT  - Journal of bone and mineral research : the official journal of the American 
      Society for Bone and Mineral Research
JID - 8610640
RN  - 0 (DNA Primers)
RN  - 0 (Parathyroid Hormone-Related Protein)
RN  - 0 (Proteins)
RN  - M801H13NRU (Azacitidine)
SB  - IM
MH  - Animals
MH  - Azacitidine/pharmacology
MH  - Base Sequence
MH  - Bone and Bones/embryology
MH  - *Calcification, Physiologic
MH  - Cattle
MH  - Cells, Cultured
MH  - Chondrocytes/*cytology/drug effects
MH  - DNA Primers
MH  - Immunohistochemistry
MH  - *Models, Biological
MH  - Parathyroid Hormone-Related Protein
MH  - Phenotype
MH  - Proteins/pharmacology
EDAT- 2001/02/24 12:00
MHDA- 2001/04/21 10:01
CRDT- 2001/02/24 12:00
PHST- 2001/02/24 12:00 [pubmed]
PHST- 2001/04/21 10:01 [medline]
PHST- 2001/02/24 12:00 [entrez]
AID - 10.1359/jbmr.2001.16.2.309 [doi]
PST - ppublish
SO  - J Bone Miner Res. 2001 Feb;16(2):309-18. doi: 10.1359/jbmr.2001.16.2.309.