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마우스 거핵구집락형성 및 분화에 대한 Interleukin-3의 효과
백남희,김현태,박애자 중앙대학교 의과대학 의과학연구소 1992 中央醫大誌 Vol.17 No.3
Interleukin 3(IL-3), a hematopoietic colony stimulating factor, promotes the proliferation of multiple murine hematopoietic lineage, influences on the megakaryocytopoiesis. Megakaryocyte has been known to require two factors for colony formation ; an inducer of clonal growth of megakaryocyte progenitors(megakaryocyte colony-stimulating factor ; Meg-CSF) and a promotor of maturation and differentiation(megakaryocyte potentiator ; Meg-potentiator). However, the effect of IL-3 in regard with its role as Meg-CSF and/or Meg-potentiator on the megakaryocytopoiesis has not been well understood yet. The total bone marrow and megakaryocyte colony count were performed and megakaryocyte precursor ploidity was measured by flow cytometry(FACScan, Becton-Dickinson) to evaluate the effect of IL-3 with variable concentations on murine hematopoiesis, especially megakaryocytopoiesis in vitro liquid culture system. The results were obtained as follows; 1) Total colony in control group was 106.45±83.493×10^3/mL per 7×10^6 bone marrow(BM) cells peak at day 2 in culture, and decreased gradually to be 54.417±43.105×10^3mL at day 4,881.200±49.106×10^3mL at day 6. 2) In a group treated with 500U/mL of IL-3, total colony count was highest with 222.231±72.784×10^3/mL at day 2 and 175.910±97.290×10^3/mL at day 4, were 2-3 times higher than that in control gruop. However it was 59.167±31.254×10^3mL at day 6, without showing any significant difference in comparison with that in control group. 3) The ratio of acetylcholinesterase positive megakaryocyte colonies to total cultured colonies was 22.0±19.6% in IL-3 treated group, which was not statistically different from that of the control group(16.9±6.9%). 4) The total colony counts was 1 to 1.4 times in 100U/mL and 1.6 to 2.0 times higher in 250U/mL concentrations of IL-3 than that in control group showing dose dependence in its increment. But it was only 1.3 to 1.5 times higher in 500U/mL, and was lower than that in 250U/mL of IL-3 suggesting inhibitory effect in higher concentration. 5) The distributions of ploidity in megakaryocyte colony cells in IL-3 treated group were 2N 59.9-72.7%, 4N 11.0-16.1%, 8N 5.7-11.0%, 16N 6.9-15.1%, 32N 1.7-8.1%, and were 2N 50.8-64.0%, 4N 11.0-16.0%, 8N 11.7-18.0%, 16N 8.6-15.2%, 32N 6.4-9.3% in control group. There was no significant difference in plodity between the two groups. And there was no change of ploidity during 2 to 6 days of culture period in both group. These findings suggested that IL-3 acts on multi-lineage committed, especially primitive stem cells, independent of lineage commitment. The IL-3 stimulates the early mitotic stage of megakaryocytopoiesis, and does not affect the endomitosis. Its stimulation is dose-dependent both at the level of 100 and 250U/mL but it is inhibitory at the level of 500U/mL. And it is concluded that Il-3 increase the numbers of the megakaryocyte colony without affecting ploidization.