凝血因子Ⅸ基因敲除对小鼠骨髓间充质干细胞生物学特性无直接影响

Biological characteristics of bone marrow mesenchymal stem cells derived from coagulation factor Ⅸ knockout mice

背景:凝血因子Ⅸ基因敲除有可能通过影响造血系统的稳定性从而影响骨髓造血微环境中骨髓间充质干细胞的生物学特性目的:鉴定凝血因子Ⅸ基因敲除C57BL/6J-F9tm1.Dws小鼠骨髓间充质干细胞的生物学特征。方法:采用改进的全骨髓贴壁细胞分离法分离培养凝血因子Ⅸ基因敲除C57BL/6J-F9tm1.Dws小鼠骨髓间充质干细胞,流细胞术鉴定其表面标记的表达,以特殊诱导培养基诱导骨髓间充质干细胞干向成骨细胞,脂肪细胞,软骨细胞方向分化。结果与结论:分离的骨髓间充质干细胞形态均一为梭形,增殖能力和自我更新能力强。流式细胞术检测细胞表面标识CD44CD56、CD73、CD90、CD105、CD106、CD166及CD271表达呈阳性,CD34、CD11b表达呈阴性。分离的骨髓间充干细胞具有向成骨细胞,脂肪细胞,软骨细胞方向分化的能力。说明凝血因子Ⅸ基因敲除C57BL/6J-F9tm1.Dws小鼠的骨间充质干细胞仍具备良好的干细胞的生物学特性,初步确定,凝血因子Ⅸ基因敲除对骨髓间充质干细胞的生物学特性并直接影响。

BACKGROUND：Coagulation factor Ⅸ knockout may affect the biological characteristics of bone marrow mesenchymal stem cells（BMSCs） of the bone marrow microenvironment by affecting the hematopoietic system stability.OBJECTIVE：To characterize the biological properties of BMSCs derived from coagulation factor Ⅸ knockout mice.METHODS：BMSCs were isolated from coagulation factor Ⅸ knockout mice C57BL/6J-F9 tm1.Dws by using a modified adherent culture method.The cells were detected for their expression of general MSCs surface markers by using flow cytometry.The functionality was assessed for their multipotency to differentiate into osteoblasts,adipocytes and chondrocytes upon certain induction conditions.RESULTS AND DISCUSSION：The BMSCs showed spindle-shape morphology and high capacity for proliferation and self-renewal.The cells were positive for CD44,CD56,CD73,CD90,CD105,CD106,CD166 and CD271,but negative for CD34 and CD11b.They demonstrated the ability to differentiate into osteoblasts,adipocytes and chondrocytes.These results illustrate the BMSCs derived from coagulation factor Ⅸ knockout mice C57BL /6J-F9 tm1.Dws have perfect stem cell biological characteristics.The results initially suggest that deficiency of coagulation factor Ⅸ has no effect on the biological characteristics of BMSCs.