CAJ | 학술논문

目的制备角质生长因子( KGF)与缺氧诱导因子( HIF)修饰的大鼠骨髓间充质干细胞( MSCs) ,评价细胞因子表达效率及其活性. 方法骨髓全细胞贴壁培养法分离培养大鼠骨髓MSCs,经免疫荧光法检测细胞表面标志物后,以KGF、HIF重组腺病毒感染,酶联免疫吸附试验( ELISA)检测KGF、HIF的表达水平,成骨细胞增殖实验评价KGF、HIF的活性.结果获得了可在体外连续传代、高表达CD44、CD133表面分子的骨髓MSCs;KGF、HIF细胞因子获得稳定表达,且在感染24 h即出现表达,随培养时间的延长表达量逐渐上升,至120 h开始下降;骨髓MSCs培养上清含量为50% ~100%时成骨细胞增殖水平明显提高.结论成功制备可稳定表达KGF、HIF的大鼠骨髓MSCs,其表达的KGF、HIF因子含量高,可促进成骨细胞的增殖,具有良好的生物活性.
목적 제비각질생장인자( KGF)여결양유도인자( HIF)수식적대서골수간충질간세포( MSCs) ,평개세포인자표체효솔급기활성. 방법 골수전세포첩벽배양법분리배양대서골수MSCs,경면역형광법검측세포표면표지물후,이KGF、HIF중조선병독감염,매련면역흡부시험( ELISA)검측KGF、HIF적표체수평,성골세포증식실험평개KGF、HIF적활성.결과 획득료가재체외련속전대、고표체CD44、CD133표면분자적골수MSCs;KGF、HIF세포인자획득은정표체,차재감염24 h즉출현표체,수배양시간적연장표체량축점상승,지120 h개시하강;골수MSCs배양상청함량위50% ~100%시성골세포증식수평명현제고.결론 성공제비가은정표체KGF、HIF적대서골수MSCs,기표체적KGF、HIF인자함량고,가촉진성골세포적증식,구유량호적생물활성.
Objective To prepare bone marrow mesenchymal stem cells (MSCs) modulated by kinetics gain factor (KGF) and hypoxia-inducible factor (HIF) in rats, and to evaluate the expression efficiencies and activities of KGF and HIF. Methods Bone marrow mesenchymal stem cells of rats were isolated and cultured using whole bone mar-row cells adherence method, and cell surface markers were identified using immunofluorescence testing, and then adeno-virus infections were restructured by KGF and HIF. The expression levels of KGF and HIF were detected using enzyme linked immunosorbent assay ( ELISA) , and the activities of KGF and HIF were evaluated using proliferation assay of oste-oblasts. Results Serial passage and high expressions in vitro of surface markers CD44 and CD133 were obtained in bone marrow mesenchymal stem cells, and the expressions of CD44 and CD133 were stable. The CD44 and CD133 were ex-pressed at the 24th h infection, and the expression levels were increasing with prolonged time, and the levels began to de-crease at the 120th h infection. The osteoblasts proliferation was obvious when supernatant content was 50%-100% by bone marrow MSCs culture. Conclusion KGF/HIF modulated rat bone marrow MSCs can be successfully achieved in rats. KGF and HIF can be highly expressed in these cells, which may promote osteoblastic multiplication with good bio-logical activity.