Primary culture and proliferation activity identification of rabbits subchondral bone cell / 国际生物医学工程杂志

ABSTRACT

ObjectiveTo study the method of cell isolation,primary culture and identification of subchondral bone cell of osteoarthritis(OA) rabbits.Methods The rabbit instable knee joint models were made by modified Hulth modeling method.The osteoblasts were harvested from the subchondral bone of rabbits by collagenase and tissue explants attachment.The morphology observation and biological identification were performed by inverted microscope and immunocytochemistry staining,respectively.The proliferative activity of cells were detected by MTT and the expression of Ⅰ-collagen at gene level was detected.ResultsThe cells started to appeared on the 11th day after the attachment.The cells form were fusiformis and triangle,the nucleolus were clear.The cultured cells had typical osteoblast morphological characteristics.The cells obtained from subchondral bone of rabbits were identified to be osteoblast by immunocytochemistry staining.The proliferative activity of cells were equably proliferation which detected by MTT.ConclusionThe modified method provides better way to obtain ideal subchondral osteoblast and the co-culture method is suitable for the study of OA microenvironment,which can simulate interactions of the subchondral osteoblast,synovial cells and chondrocyte.