携带α-地中海贫血基因人胚胎干细胞系的建立  

Establishment of Human Embryonic Stem Cell Lines Carrying with α-Thalasimia Gene

作　　者：麦庆云[1] 邓捷[1] 彭文林[1] 庄广伦[1] 周灿权[1] 

机构地区：[1]中山大学附属第一医院生殖中心,广东广州510080

出　　处：《中山大学学报（医学科学版）》2009年第5期551-555,576,共6页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(30801239);广东省自然科学基金(07301429);教育部博士点新青年教师基金(200805581164)

摘　　要：【目的】建立携带α-地中海贫血纯合子基因人胚胎干细胞系。【方法】收集经胚胎种植前遗传学诊断技术诊断为携带α-地中海贫血纯合子基因的囊胚,机械法分离内细胞团,用无血清培养基进行人胚胎干细胞培养,建立携带α-地中海贫血纯合子基因的人胚胎干细胞系。对所得人胚胎干细胞进行α-地中海贫血基因的DNA序列分析与全能性鉴定。【结果】本研究共收集44枚PGD筛查后囊胚,分离12个囊胚内细胞团,建立两株人胚胎干细胞系。两株人胚胎干细胞系均携带α-地中海贫血纯合子基因。两株胚胎干细胞系均有正常的染色体核型、OCT-4基因与细胞表面抗原SSEA-3、SSEA-4、TRA-1-60与TRA-1-80的表达,能够向三胚层细胞分化。【结论】携带α-地中海贫血纯合子基因的人胚胎干细胞具有向三胚层细胞分化的全能性,携带致病基因的胚胎可用于建立携带遗传疾病基因的人胚胎干细胞系。【Objective】 To establish human embryonic stem cell lines carrying with α-thalasimia gene. 【Methods】 α- Thalasimia homozygote affected blastocysts were collected from preimplantation genetic diagnosis （PGD） treatment cycles and the inner cell masses were isolated by mechanical method. Serum-free culture medium was used in the cultivation of human embryonic stem cells. α-thalasimia genes of two embryonic stem cell lines were confirmed by DNA sequence analysis. The pluripotency of two embryonic stem cell lines were identified. 【Results】 In this study, we collected 44 blastocysts carrying with α-thalasimia gene and established two embryonic stem cell lines. Both embryonic stem cell lines are α-thalasimia homozygote affected and retained the property of pluripotent cells. Two cells lines had normal karyotypes and expressed transcription factor OCT-4, stage specific expressed antigen-4 （SSEA-4）, SSEA-3,TRA-1-60, and TRA-1-80. Both ES cell lines kept the capability to differentiate to the cells and tissues derived from three germ layers. 【Conclusions】 Embryonic stem cells carrying with α-thalasimia gene retained pluripotency. Embryos affected with disease gene can be used for isolation of human embryonic stem cell lines carrying with inherited disease gene.

关 键 词：人类胚胎干细胞 Α-地中海贫血 多能性 

分 类 号：R321-33[医药卫生—人体解剖和组织胚胎学]