We have developed a in vitro culture system of osteoblasts isolated from juvenile rabbit periosteum. The cultivation could be served as an osteoblast biological model. The newly developed method has the following advantages: (a) The periosteum is easy to obtain. (b) The periosteal cambium layer contains a great deal of osteoblasts. (c) The procedure of isolating osteoblasts from periosteum is less complex than that from bone tissue. The cultivation could be regarded as a useful technique for studying the metabolism and osteogenic capacity of osteoblasts in vitro.