小鼠诱导性多能干细胞诱导成骨分化的初步研究

›› 2015, Vol. 6 ›› Issue (4): 193-197.

小鼠诱导性多能干细胞诱导成骨分化的初步研究

钱浩¹,徐增琦²,李杨¹,徐镭¹,程杰¹,江宏兵³,吴煜农⁴,Jake Chen⁵,叶金海²

1. 南京医科大学附属口腔医院口腔颌面外科
2. 南京医科大学附属口腔医院颌面外科
3. 南京医科大学口腔医学院
4. 南京医科大学附属口腔医院
5. 美国塔夫茨大学牙学院口腔生物学研究室

收稿日期:2015-09-25 修回日期:2015-11-22 出版日期:2015-12-25 发布日期:2015-12-30
通讯作者: 叶金海 E-mail:yjh98001@163.com
基金资助:
国家自然科学基金项目;江苏省自然科学基金;江苏高校优势学科建设工程

Research of induced pluripotent stem cells of mouse to induce osteogenic differentiation

Received:2015-09-25 Revised:2015-11-22 Online:2015-12-25 Published:2015-12-30

摘要/Abstract

摘要： 目的：研究小鼠诱导性多能干细胞（induced pluripotent stem cells，iPSCs）向成骨细胞分化的能力，并观察Osterix对iPSCs成骨分化能力的影响。方法：利用体外细胞培养和病毒转染的方法，结合定量RT-PCR以及组织化学的方法检测在成骨诱导的条件下，iPSCs中成骨相关基因、蛋白的表达变化。结果：小鼠iPSCs经悬滴培养可以形成拟胚体，在成骨诱导条件下可以向成骨细胞分化。病毒转染并不影响iPSCs的多向分化潜能；与对照组相比，转染Osterix的iPSCs形成的矿化结节、碱性磷酸酶活性、成骨相关基因的表达均有明显增加（P<0.05）。结论：在成骨诱导液培养条件下，小鼠iPSCs可以作为种子细胞向成骨细胞分化，并且过表达转录因子Osterix可以进一步促进iPSCs的成骨分化。

Abstract: Objective：To characterize and observe the effect of Osterix on the osteogenic differentiation of mouse iPSCs. Methods： Through iPS cell culture, gene transfection, semi-quantitative RT-PCR and histology, we analyzed the alteration of osteogenesis related genes and protein in the mouse iPSCs before and after Osterix transfection. Results：Suspension of iPS cells in differentiation culture (drop) can form embryoid bodies (EB). After the plasmid transfection, the potential of multi-directional differentiation of iPS cells was not found to have changed. Moreover, compared with the control group, the activity of ALP in the formation of mineralized nodules, osteogenesis related genes (Runx2, bone sialoprotein and osteocalcin) expression were significantly enhanced in the iPSCs transfected with Osterix (P<0.05). Conclusions：iPSCs can be differentiated to bone cells, and the transcription factor Osterix can promote the osteogenic differentiation of iPS cells.

钱浩徐增琦李杨徐镭程杰江宏兵吴煜农 Jake Chen 叶金海. 小鼠诱导性多能干细胞诱导成骨分化的初步研究[J]. , 2015, 6(4): 193-197.

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