CAJ | 학술논문

目的研究 CD226在造血干 /祖细胞上的表达分布。 TPO诱导胎肝造血干 /祖细胞增殖分化过程中， CD226表达的变化及可能的信号调控途径。方法采用鸡尾酒单抗阴性分离富集 Lin－造血干 /祖细胞，培养于含 TPO的无血清培养体系中，用 PD98059阻断 MAPK信号转导途径，流式细胞仪双荧光分析 CD226、 CD34、 CD41a和 CD61的表达。结果 CD226在 Lin－ CD34＋细胞和 Lin－ CD34－细胞上均有表达。 TPO可诱导 Lin－ CD41a－ CD226－细胞先分化为 CD41a＋ CD226－细胞，再进一步分化为 CD41a＋ CD226＋细胞， TPO也可诱导 Lin－ CD226＋细胞表达 CD41a和 CD61。在培养的早期， PD98059可抑制 CD34＋ CD226＋细胞的减少，在晚期则可抑制 CD41a＋ CD226＋细胞的增多。结论 CD226与造血干 /祖细胞和巨核细胞的增殖分化可能具有密切的关系。
목적연구 CD226재조혈간 /조세포상적표체분포。 TPO유도태간조혈간 /조세포증식분화과정중， CD226표체적변화급가능적신호조공도경。방법채용계미주단항음성분리부집 Lin－조혈간 /조세포，배양우함 TPO적무혈청배양체계중，용 PD98059조단 MAPK신호전도도경，류식세포의쌍형광분석 CD226、 CD34、 CD41a화 CD61적표체。결과 CD226재 Lin－ CD34＋세포화 Lin－ CD34－세포상균유표체。 TPO가유도 Lin－ CD41a－ CD226－세포선분화위 CD41a＋ CD226－세포，재진일보분화위 CD41a＋ CD226＋세포， TPO야가유도 Lin－ CD226＋세포표체 CD41a화 CD61。재배양적조기， PD98059가억제 CD34＋ CD226＋세포적감소，재만기칙가억제 CD41a＋ CD226＋세포적증다。결론 CD226여조혈간 /조세포화거핵세포적증식분화가능구유밀절적관계。
Aim To investigate the expression of CD226 on Lin－ hematopoietic stem/progenitor cells and TPO－ induced Lin－ cells as well as possible pathway of signal transduction for regulating expression of CD226 by TPO. Methods Lin－ hematopoietic stem/progenitor cells isolated from fetal liver mononuclear cells by negative selection with human progenitor enrichment－ antibodies cocktail including anti CD2,  CD3,  CD14,  CD16,  CD19,  CD24,  CD56,  CD66b mAbs and glycophorin A mAb were incubated in serum－ free IMDM supplemented with 15％ of mixture of BSA, insulin, transferrin (BIT9500) and 100μ g/L TPO for 18 days. In some experiments, PD98059, a MEK1 inhibitor, was added at a concentration of 20μ mol/L. The expressions of CD226, CD34, CD41a and CD61 were analyzed by using flow cytometry with double－ labelling technique. Results CD226 was expressed on both Lin－ CD34＋ cells and Lin－ CD34－ cells. TPO firstly induced Lin－ CD41a－ CD226－ cells to differentiate into CD226－ CD41a＋ /CD61＋ cells and then into CD226＋ CD41a＋ /CD61＋ cells. TPO could also induce Lin－ CD226＋ cells to express CD41a/CD61. PD98095 could suppress decrease in CD34＋ CD226＋ cells in the early period of the Lin－ cell culture and inhibit increment in CD41a＋ CD226＋ cells in the later period. Conclusion CD226 seems to have a close relation with proliferation and differentiation of hematopoietic stem/progenitor cells and megakaryocytic progenitor cells.