Primary culture of osteoblasts from fetal rat

Objective To establish a simple,quantities,purity proposal for osteoblast primary culture,and to lay the foundation for subsequent experimental studies. Methods From March to September 2009,at the biological laboratory of Institute of Metal Research,Chinese Academy of Science,the skull was taken from 4 fetal rats which were born within 24 hours. Use trypsin and collagenaseⅠin turn to digest osteoblasts. Using ALP staining,collagen Ⅰ immunofluorescence staining and alizarin red staining to identify the cultured cells. Results The cultured cells had typical morphological characteristics of osteoblasts;ALP staining,collagen Ⅰimmunofluorescence staining and alizarin red staining were positive. Conclusion Osteoblasts achieved by this method are pure with large quantities,so this method can be used as a reliable and efficient way of primary culture.