[In vitro culture of human osteoblasts].
OBJECTIVE
To modify the digestion and culture of human osteoblasts for releasing a large amount of cells.


METHODS
Bone tissue was harvested and digested by trypsin for several times to remove blood cells and fibroblasts. After cultured for a couple of days in incubator, bone chips were digested again by trypsin for several times, and a large amount of pure osteoblasts were released and used for researches. The remaining chips were digested by trypsin repeatedly until enough cells were harvested. Black nodes were seen in confluent cell layer. Osteoblasts were confirmed by their unique secretion of a large amount of alkaline phosphatase (AKP), osteocalcin (OCN) and type I collagen.


RESULTS
Under fluorescent microscope, black node was "golden yellow" in color, a characteristic of bone node. Secretion of AKP (2.76 +/- 0.56) IU/ml and OCN (1.875 +/- 0.549) ng/ml of osteoblasts was higher than those of fibroblasts. There was only type I collagen in supernatant without type III.


CONCLUSIONS
The modified osteoblast digestion and culture can release a large amount of pure osteoblasts in a short period with no fibroblastic adulteration.
Zhang
X
X
Shanghai Ruijin Hospital, Shanghai Institute of Traumatology and Orthopaedics, Shanghai 200025, China.
Yang
Q
Q
Deng
L
L
Zhu
Y
Y
D000469:Alkaline Phosphatase / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D006801:Humans; D010006:Osteoblasts / Q000378:metabolism / Q000502:physiology*; D015675:Osteocalcin / Q000378:metabolism
D015675:Osteocalcin; D003094:Collagen; D000469:Alkaline Phosphatase