|Budget Amount *help
¥2,100,000 (Direct Cost : ¥2,100,000)
Fiscal Year 1994 : ¥800,000 (Direct Cost : ¥800,000)
Fiscal Year 1993 : ¥1,300,000 (Direct Cost : ¥1,300,000)
1. Rat megakaryocytes (MK) were isolated by an immunomagnetic beads method as described in the report of last year. Then, IL-6 and IL-11 were compared for PPF activity because they are known to be pluripotent factors having many similarities in biological aspects although they are categolized as a MK-potentiator (Meg-POT). As a result of it, IL-11 was wuite different in the PPF activity from IL-6 because IL-11 alone is ineffective at 0.01 to 100ng/ml for the PPF activity and not synergistic to erythropoietin (Epo), and slightly effective in the coculture system of MK and stromal cells (fibroblasts), while IL-6 was remarkably effective in all cases.
2. A novel factor, a ligand for c-Mpl, is thought to be thrombopoietin (TPO). TPO delivered from Kirin Brewery Co. Ltd. was compared with IL-6 and Epo for the PPF activity in the same way. In serum-free PPF assay, TPO was strongly (100-1,000 fold) effective because the effective doses of TPO ranged from 0.00001 to 1ng/ml and effective reange was expanded up to 100ng/ml in the coculture system using porcine endothelial cells, in which the PPF activity (score of PPF) of TPO as well as IL-6 and Epo was greatly enhanced. The effective doses of IL-6 and Epo are from 0.1 to 100ng/ml and 0.01 to 10U.ml respectively.