通过患者来源iPS细胞建立阵挛性癫痫伴破碎红纤维综合症心肌细胞疾病模型  

Construction of model for cardiomyocyte disease MERRF using patient-derived iPSC

作　　者：徐天慧[1] 林颖[1] 梁栋[1] 王霞[1] 胡平[1] 许争峰[1] XU Tianhui , L1N Ying , LIANG Dong , WANG Xia , HU Ping, XU Zhengfeng(Prenatal Diagnosis Center, Obstetrics and Gynecology Hospital Affiliated to Nanjing Medical University, Nanfing 210004, Jiangsu, China)

机构地区：[1]南京医科大学附属妇产医院产前诊断中心,南京210004

出　　处：《临床检验杂志》2018年第2期90-93,共4页Chinese Journal of Clinical Laboratory Science

基　　金：江苏省自然科学基金(BK20150081)

摘　　要：目的对阵挛性癫痫伴破碎红纤维综合症(MERRF)患者来源的诱导多能干细胞(i PSC)进行体外分化,获得携带MERRF疾病相关线粒体突变的心肌细胞,评估其在建立MERRF的心肌细胞疾病模型中的价值。方法采用定向分化的方法体外将患者来源的i PSC及对照H9分化为心肌细胞,免疫荧光染色和RT-PCR对两组心肌细胞进行鉴定,并对心肌细胞的搏动频率进行统计,比较两组心肌细胞的功能。结果患者来源的i PSC及H9均成功分化为心肌细胞;在心肌分化的第10、13、15、16天,MERRF-i CMs平均搏动频率分别为13次/分、24次/分、15次/分、18次/分,H9-i CMs平均搏动频率分别为80次/分、96次/分、120次/分、120次/分,差异均具有统计学意义(P均<0.05)。结论通过患者来源i PSC向心肌细胞的分化及功能评估,获得了携带MERRF疾病相关线粒体突变的心肌细胞模型。Objective To obtain myoclonic epilepsy with ragged-red fibers（ MERRF）-specific cardiomyocytes by the differentiation in vitro of inducing pluripotent stem cells（ i PSC） derived from a MERRF patient and evaluate the application of the prepared cardiomyocytes in construction of MERRF syndrome model. Methods The patient-derived i PSCs and H9 embryonic stem（ ES） cells,the control cell line,were unidirectionally differentiated into cardiomyocytes n in vitro. The obtained cardiomyocytes were identified and validated by detecting the presence of cardiomyocyte-specific markers using immunofluorescence staining and RT-PCR. The beating frequencies were recorded to compare the functional evaluation for the two groups of cardiomyocyte. Results Both the patient-derived i PSC and H9 ES cells were differentiated into cardiomyocytes successfully. The average beating frequencies of MERRF-induced cardiomyocytes（ i CMs） were 13,24,15 and 18 times/min on the day 10,13,15 and 16 during the cell differentiation process. The average beating frequencies of H9-i CMs were 80,96,120 and 120 times/min,respectively. The beating ability of i PSC-differentiated cardiomyocytes was significantly lower than that of corresponding control（ all P〈0. 05）. Conclusion The patient-derived i PSCs may differentiated into cardiomyocytes. Based on the functional evaluation for these cardiomyocytes,the model for MERRF syndrome with mitochondrial mutations was generated and characterized in vitro.

关 键 词：阵挛性癫痫伴破碎红纤维综合症 诱导多能干细胞 线粒体遗传病 心肌分化 

分 类 号：R742.1[医药卫生—神经病学与精神病学]