|Budget Amount *help
¥2,100,000 (Direct Cost : ¥2,100,000)
Fiscal Year 1990 : ¥900,000 (Direct Cost : ¥900,000)
Fiscal Year 1989 : ¥1,200,000 (Direct Cost : ¥1,200,000)
1. Although another megakaryocytic cell line like T-33 could not be established, in our laboratory we succeeded in establishing an interesting new human myelogenous cell line derived from polycythemia vera, and an human mesothelial cell line derived from malignant mesothelioma with secondary thrombocytosis. The latter has been found to contain IL-6 and IL-3.
2. When recombinant interleukin 6 (rIL-6) as a megakaryocytic differentiation factor was used to induce megakaryocytic maturation of T-33 cells, rIL-6 alone increased intracytoplasmic platelet glycoprotein (GP IIb IIIa), and also showed synergistic effect on the appearance of GP IIb IIIa on the cell surface. This indicated that rIL-6 was effective for the megakaryocytic differentiation of the cell line too.
3. We continued the c-yes tyrosine protein kinase (TPK) assay for the megakaryocytic differentiation of T-33 cells, using the in vitro immuno-precipitation method. As a result of it, T-33 replatedly showed the dcereased c-yes TPK activity at 3 to 40 hours after the TPA treatment. Similarly, K562 showed the decrease at 16 to 40 hours after the same treatment. On the other hand, the monocytic cell line U937 showed the increase at 6 to 48 hours, and HL-60 cell line showed the decrease in the early phase and the subsequent increase at 36 hours. In the case of incunbation with IL-6, the TPK activity of T-33 increased at 40 hours. Contrary to the c-yes, the TPK activity of scr generally increased at 24 to 48 hours in all cell lines tested.