CAJ | 학술논문

背景:细胞形态结构和功能密不可分,但目前对人脐带间充质干细胞超微结构的报道还很少.目的:探讨人脐带间充质干细胞的生物学特性与原子力显微镜下超微结构的关系.方法:采用酶消化法体外分离培养并扩增人脐带间充质干细胞,取第3代细胞,用原子力显微镜在接触模式下观察成像,通过流式细胞仪检测细胞免疫表型和细胞周期,油红O染色及碱性磷酸酶染色鉴定其成脂、成骨分化潜能.结果与结论:第3代人脐带间充质干细胞强表达CD44,CD29,低表达CD106,不表达CD34;有80%以上的细胞处在G_0/G_1期,增殖指数为19.9%;成脂诱导后,油红O染色可见胞浆中有大量桔红色的小脂滴;成骨诱导后,碱性磷酸酶染色可见立方形、多边形细胞的胞质染成棕褐色.原子力显微镜下人脐带间充质干细胞以长梭形为主,细胞骨架丝明显,并形成网状连接,这与其强大的增殖、迁移、分化功能相适应.
배경:세포형태결구화공능밀불가분,단목전대인제대간충질간세포초미결구적보도환흔소.목적:탐토인제대간충질간세포적생물학특성여원자력현미경하초미결구적관계.방법:채용매소화법체외분리배양병확증인제대간충질간세포,취제3대세포,용원자력현미경재접촉모식하관찰성상,통과류식세포의검측세포면역표형화세포주기,유홍O염색급감성린산매염색감정기성지、성골분화잠능.결과여결론:제3대인제대간충질간세포강표체CD44,CD29,저표체CD106,불표체CD34;유80%이상적세포처재G_0/G_1기,증식지수위19.9%;성지유도후,유홍O염색가견포장중유대량길홍색적소지적;성골유도후,감성린산매염색가견립방형、다변형세포적포질염성종갈색.원자력현미경하인제대간충질간세포이장사형위주,세포골가사명현,병형성망상련접,저여기강대적증식、천이、분화공능상괄응.
BACKGROUND: The relationship between cellular morphosis and function is undetachable, but there is little investigation about ultrastructure of human umbilical cord mesenchymal stem cells (hUCMSCs).OBJECTIVE: To study the relationship between the functions of hUCMSCs and their ultrastructure obtained by atomic force microscope (AFM). METHODS: hUCMSCs were isolated, cultured, expanded after enzyme digestion. P3 cells were observed under AFM. Immunophenotype and cell cycle were analyzed by flow cytometry, as well as induction of the adipogenic, osteogenic differentiation of hUCMSCs were identified using Oil red O staining and alkaline phosphatase staining. RESULTS AND CONCLUSION: hUCMSCs at passage 3 were strongly positive for CD44 and CD29, weakly positive for CD106, but negative for hematopoietic marker CD34. Cells in G_0/G_1 phase accounted for 80%. Proliferation index was 19.9%. Following adipogenic induction, alkaline phosphatase staining demonstrated brown cytoplasm in cube and polygonal cells. AFM showed hUCMSCs were spindle shape, obvious cytoskeletal filament that connected into nets, which fit for the strong capacities for proliferation, migration and differentiation.