建立非基因整合原发性骨髓纤维化诱导性多能干细胞株  

Establishment of Integration-Free Human Induced Pluripotent Stem Cells from A Patient with Primary Myelofibrosis

作　　者：许静[1] 盛梦瑶 周圆[1] 邢文[1] 白洁[1] 温伟[1] 纪光臻[1] 张鸿雁[1] 金晖[1] 吕翠翠[1] 袁卫平[1] 张孝兵[1] 程涛[1] 

机构地区：[1]中国医学科学院北京协和医学院,血液病医院血液学研究所,实验血液学国家重点实验室,天津300020

出　　处：《中国实验血液学杂志》2015年第5期1415-1421,共7页Journal of Experimental Hematology

基　　金：科技部重大基础研究项目(2012CB966601;2013CB9669022015CB964902);国家自然科学基金(81170512);天津市应用基础研究重点项目(13JCZDJC31200)

摘　　要：目的:利用诱导多能性干细胞(iPSC)技术建立原发性骨髓纤维化(PMF)诱导多能性干细胞系,为研究血液病的发生发展提供一个实验模型。方法:采用非基因整合型质粒重编程携带JAK2V617F突变的PMF患者骨髓来源的单个核细胞,诱导生成该患者特异的iPS细胞株。结果:采用此方法建立的iPS细胞株,在体外能够稳定传代,无外源性基因整合,多能性基因表达水平与人胚胎干细胞类似,在体内具有形成三胚层结构的能力。测序结果显示,所建立的患者的iPS细胞株携带不同负荷的JAK2V617F突变。结论:成功地建立非基因整合的PMF患者特异的iPS细胞株,这为研究PMF的发病机制、化疗药物筛选及实现精准化治疗提供了一个重要的疾病模型。Objective: To establish the primary myelofibrosis( PMF)-induced pluripotent stem cell line( iPSC) by means of iPSC techinique so as to provide a experimental model for studing the blood disease mechanisms. Methods:Induced pluripotent stem cells were generated from mononuclear cells isolated from a PMF patient with JAK2V617 F mutation by using episomal vectors. Results: PMF- derived iPSC was established from the paitent with JAK2V617 F gene mutation.The PMF-iPSC could be stably passaged,highly expressed pluripotent genes as human embryonic stem( ES)cells,and were able to form teratoma in NOD /SCID mice in vivo. H ﹠ E staining of the teratoma showed the presence of tissue type derived from all three embryonic germ layers. Sanger sequencing confirmed that PMF-derived iPSC carried different allele burdens of JAK2V617 F gene mutation. Conclusion: The interation-free iPSC from primary myelofibrosis patient in vitro has been established. This PMF-derived iPSC line provides a valuable tool for studying the pathogenesis,screening of chimical drugs and realizing the standard therapy of PMF.

关 键 词：诱导多能性干细胞 原发性骨髓纤维化 非整合 Episomal载体 

分 类 号：R551.3[医药卫生—血液循环系统疾病]