Apoptosis of rat osteoblasts in process of calcification in vitro.
OBJECTIVE
To establish a cell model of osteoblasts retaining their differentiated phenotype in culture and observe the apoptosis of osteoblasts in the process of calcification using a novel acetylcholinesterase (AChE) staining method.


METHODS
Osteoblasts were isolated enzymatically from skull of newborn SD rats; alkaline phosphatase (AKP) activity was determined by reformed cobalt method and azo dye method; mineral deposition was assessed with Von Kossa staining and Fluo-3 staining; a novel AChE staining method was used to assay cellular apoptosis based on the higher expression of AChE in apoptotic cells.


RESULTS
During the 44 d of cells cultured, primary rat skull-derived osteoblasts progressively developed into a bone-like tissue of multi-layered nodules of cells with mineralized extra-cellular matrix and the apoptotic cells increased while the matrix calcificated.


CONCLUSIONS
The phenotype of developmental sequence of rat skull-derived osteoblasts can reflect the maturation of osteoblasts in vitro. It is a convenient model for the research of osteoblasts biology.
Gu
Qi
Q
Shanghai Institute of Gerontology, Huadong Hospital, Shanghai 200040, China. gu_qi00@hotmail.com.
Zhu
Han-Min
HM
Zhang
Xue-Jun
XJ
D000469:Alkaline Phosphatase / Q000378:metabolism*; D000818:Animals; D000831:Animals, Newborn; D017209:Apoptosis / Q000502:physiology*; D002113:Calcification, Physiologic / Q000502:physiology*; D002478:Cells, Cultured; D010006:Osteoblasts / Q000166:cytology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012886:Skull / Q000166:cytology
D000469:Alkaline Phosphatase