This study was performed to determine whether the growth rate and metastatic potential of tumors generated by spontaneous lung metastases is influenced by transplantation methods. Three different tumors syngeneic to C3Hf/Kam mice were studied: the SA-NH and SA-4020 sarcomas and the hepatocarcinoma HCA-I. Solitary tumors in the legs of mice were generated by a single metastatic nodule taken at random from lung metastases, by a single metastatic nodule taken from each mouse with the highest number of metastases, by a mixture of cells from lung metastases taken randomly, or by a mixture of cells from primary leg tumors. These transplantation procedures were repeated for two to four isotransplant generations. Repeated isotransplants of primary tumors showed little if any change in the growth rate and metastatic spread. In contrast, primary tumors derived from spontaneous metastases frequently exhibited a decrease in their growth rate and an increase in metastatic potential. This was particularly frequent when tumors were established from single metastatic nodules taken randomly from the lung, or taken from lungs that contained the largest number of metastatic nodules. The magnitude of this change varied greatly among the three tumors studied. Increased metastatic formation in the lung was also frequently associated with slower growth of the primary tumors. Thus, transplantation methods used for establishing primary tumors have an important influence on the metastatic potential of tumor transplants.