Analysis of CCN4 Function in Osteogenic and Osteoclastic Cells Using Gain and Loss of Function Approaches.

Analysis of CCN4 function in bone was assessed using both gain and loss of function approaches. In mice this was done by genetic engineering and homologous recombination to create mice with complete ablation of the protein. For human skeletal cells adenoviral gene transfer and shRNA were used for gain and loss of function respectively. Here we describe procedures used to make and then analyze osteogenic and osteoclastic cells with or without CCN4 to determine its role in osteogenic differentiation.