CAJ | 학술논문

目的：探讨体外诱导小鼠骨髓CD45-细胞向窦房结起搏样细胞定向分化的方法。方法免疫磁珠分选小鼠骨髓CD45-细胞，体外培养扩增后，采用添加了5-氮胞苷和小鼠窦房结组织培养上清液的诱导培养基培养，经免疫荧光染色检测起搏样细胞标志物神经丝蛋白-160(NF-160)和超级化激活环核苷酸门控阳离子通道(HCN)4表达，并采用qRT-PCR定量检测起搏基因NF-160、HCN4和HCN2表达。结果小鼠骨髓CD45-细胞经体外诱导培养后得到的分化细胞表达NF-160和HCN4；qRT-PCR分析显示，与体外培养的小鼠窦房结细胞比较，分化起搏样细胞NF-160和HCN4的表达升高，HCN2的表达降低。结论体外培养能够将小鼠骨髓CD45-细胞诱导分化为窦房结起搏样细胞，为构建生物起搏器种子细胞提供选择。
목적：탐토체외유도소서골수CD45-세포향두방결기박양세포정향분화적방법。방법면역자주분선소서골수CD45-세포，체외배양확증후，채용첨가료5-담포감화소서두방결조직배양상청액적유도배양기배양，경면역형광염색검측기박양세포표지물신경사단백-160(NF-160)화초급화격활배핵감산문공양리자통도(HCN)4표체，병채용qRT-PCR정량검측기박기인NF-160、HCN4화HCN2표체。결과소서골수CD45-세포경체외유도배양후득도적분화세포표체NF-160화HCN4；qRT-PCR분석현시，여체외배양적소서두방결세포비교，분화기박양세포NF-160화HCN4적표체승고，HCN2적표체강저。결론체외배양능구장소서골수CD45-세포유도분화위두방결기박양세포，위구건생물기박기충자세포제공선택。
Objective To study method on inducing directional differentiation of mouse bone marrow CD45-cells into sinoatrial node pacemaker-like cells in vitro. Methods Mouse bone marrow CD45-cells were derived by using im-munomagnetic beads system. After amplification in vitro, induced medium supplemented with 5-azacytidine and culture supernatant liquid of mouse sinoatrial node cells was used to cultivate. The expression on pacemaker-like cells markers of neurofilament protein (NF-160) and hyperpolarization activated cyclic nucleotide-gated potassium channel (HCN) 4 were detected by immunofluorescence staining. The expression of pacing genes (NF-160, HCN4 and HCN2) were de-tected by qRT-PCR. Results Differentiated cells of bone marrow CD45-cells after induced by culture in v itro ex-pressed NF-160 and HCN4. qRT-PCR analysis showed, compared with cultured sinoatrial node cells in v itro, the ex-pression of NF-160 and HCN4 were higher, while expression of HCN2 was lower. Conclusion The sinoatrial node pacemaker-like cells can be induced and differentiated from mouse bone marrow CD45-cells, and the cells will be one choice of ideal seeding cells for biological pacemaker building.