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The primary culture and the identification of human fetal osteoblasts

Li Dehua, Liu Baolin, Wu Junzheng et al School of Stomatology, The Fourth Military Medical University, Xi'an 710032  
Aim:Based on the advantages and disadvantages of enzyme-disgestion technique commonly used for the isolation of osteoblasts from bone tissue, we investigated the feasibility of the explant technique and its practicable value. Methods: The bone explants were harvested from the calvaria of 6-month fetuses after legal abortion and cultured in DMEM containing 50μg/ml of L- ascordic acid. The identification of derived cells was done by histochemical staining of alkaline phosphatase( ALPase) , immunohistochemical staining of osteocalcin, von- Kossa staining of calcified nodules and the observation of cell morphology. Results: After 5 days of cell culture,some triangle-shaped or short spindle-shaped cells migrated from the explants,and at the same time,the bone explants tended to break into small pieces.After 7 days, triangle- shaped or square- shaped cells appeared under the breaking explants.And finally, the cells reached confluency after 12 days.The derived cells were identified to be osteoblastic cells by the criteria of ALPase,osteocalcin and calcifing nodules formation. Conclusions: As to the primary isolation of osteoblasts from bone,the explant technique is a reliable, efficient and effective method.
【CateGory Index】: Q343.6,
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