两种细胞来源的21-三体综合征诱导多能干细胞系的建立及比较  

Generation and Comparison of Human Down Syndrome-Induced Pluripotent Stem Cells from Two Types of Cells

作　　者：杜融[1] 骆玉梅[1] 王鼎[1] 孙筱放[1] 陈耀勇[1] 

机构地区：[1]广东省产科重大疾病重点实验室广东省高校生殖与遗传重点实验室广州医科大学附属第三医院,广州510150

出　　处：《中国细胞生物学学报》2014年第7期884-891,共8页Chinese Journal of Cell Biology

基　　金：广州市科信局重大项目(批准:2011Y1-00038)资助的课题~~

摘　　要：21-三体综合征是染色体异常导致的疾病,通过重编程21-三体综合征患儿两种组织来源的细胞成为多能干细胞,比较两种组织来源的细胞建立21-三体综合征诱导多能干细胞(T21-iPSCs)系的效率,为进一步研究21-三体综合征发病机制提供细胞模型,并为选择高效制备T21-iPSCs的组织类型提供理论依据。该实验利用慢病毒介导4种转录因子(Oct4、Sox2、Klf4、c-Myc)分别诱导人21-三体综合征的羊水细胞和胎儿皮肤成纤维细胞,建立诱导多能干细胞系(Trisomy 21 human amniotic fl uid induced pluripotent stem cells,T21 hAF-iPSCs;Trisomy 21 human dermal fi broblast induced pluripotent stem cells,T21 hDF-iPSCs),T21 hAF-iPSCs及T21 hDF-iPSCs在蛋白和mRNA水平上均表达人胚胎干细胞的多能性分子标记,如Oct4、Nanog等,具有在体外及体内分化三个胚层的能力,其在培养过程中能维持异常核型并能维持自我更新状态。结果发现,利用羊水细胞建立T21-iPSCs效率高于皮肤成纤维细胞,羊水细胞可能是制备T21-iPSCs的理想细胞类型。Down syndrome （DS）, or Trisomy 21 （T21） syndrome, one of the most common chromosomal abnormalities, is caused by an extra duplication of chromosome 21. The generation of induced pluripotent stem cells （iPSCs） is a critical step in understanding the developmental stages of complex chromosomal diseases. In this study, we have generated DS-specific induced pluripotent stem cells from human amniotic fluid-derived cells and dermal fibroblast cells through lentiviral delivery of four human transcription factors （Oct4/Sox2/Klf4/c-Myc）. The DS-iPSCs （Trisomy 21 human amniotic fluid induced pluripotent stem cells, T21 hAF-iPSCs; Trisomy 21 human dermal fibroblast induced pluripotent stem cells, T21 hDF-iPSCs） showed characteristics similar to those of human embryonic stem cells, particularly the morphology and pluripotent-specific transcription-factor （Oct4, Nanog） expression. The pluripotency of DS-iPSCs was also tested in vitro and in vivo. Embryoid bodies and teratomas were formed and showed the expression of differentiated markers for three germ layers. The DS-iPSCs showed a goodself-renewal condition and maintained the karyotypes after long-term culturing in vitro. It was found that generation of iPSCs from human amniotic fluid cells were more rapid and efficient than dermal fibroblast cells. Amniotic fluid cells may be a preferred tissue for generating T21-iPSCs.

关 键 词：21-三体综合征 羊水细胞 人皮肤成纤维细胞 诱导多能干细胞 

分 类 号：R714.5[医药卫生—妇产科学]