Establishment and characterization of a murine Gardner's osteosarcoma cell line (GOS/T): purification of bone-specific alkaline phosphatase (ALPase) and use of anti-ALPase as a diagnostic acid.

Gardner's murine osteosarcoma has been successfully cultured and maintained as a clonal cell line (GOS/T), capable of forming a tumor with neoplastic bone and osteoid tissue by inoculation of 1 x 10(7) cultured cells. Alkaline phosphatase (ALPase) level in the culture medium and the serum from the tumor-bearing mice increased significantly three times and 16 to 105 times, respectively. The bone-specific ALPase was purified by butanol extraction following gel-filtration through Sephadex G-200. The molecular weight of ALPase was determined as 420,000, which was three times 140,000, the molecular weight of a subunit. Immunofluorescence staining using anti-ALPase antiserum, which was made by inoculation of partially purified ALPase, revealed a positive reaction for GOS/T cell line and osteoblasts of the fetal mice. The data presented here suggest that the cell line of GOS/T can be regarded as an established cell line, and the immunological reaction with anti-ALPase antibody is a useful model system to study human osteosarcoma.