title: [In vitro culture of human osteoblasts].
abstract: 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.
authors.affiliation: Shanghai Ruijin Hospital, Shanghai Institute of Traumatology and Orthopaedics, Shanghai 200025, China.
authors.forename: X
authors.initials: X
authors.lastname: Zhang
authors.forename: Q
authors.initials: Q
authors.lastname: Yang
authors.forename: L
authors.initials: L
authors.lastname: Deng
authors.forename: Y
authors.initials: Y
authors.lastname: Zhu
mesh_terms: 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
chemical_list: D015675:Osteocalcin; D003094:Collagen; D000469:Alkaline Phosphatase