IPS 课件 Induced Pluripotent Stem Cells(iPSCs).ppt

造血干细胞 造血干细胞是指骨髓中的干细胞，具有自我更新能力，能发育生成各种类型的血细胞。造血干细胞可以救助很多患有血液病的人们，最常见的就是白血病。 脐带血干细胞 间充质干细胞 神经干细胞 成体干细胞 干细胞 干细胞分类 胚胎干细胞 The Third Reprogramming Strategies 2. Generation of iPS cells from Adult HDF Direct differention into Neural Cells Marker gene expression 诱导多功能干细胞（iPS） 2006年日本京都大学Yamanaka领导的实验室在世界著名学术杂志《细胞》上率先报道在小鼠皮肤成纤维细胞细胞中高表达上述四种转录因子，可诱导细胞发生转化，产生的新细胞在形态、基因和蛋白表达、表观遗传修饰状态、细胞倍增能力、类胚体和畸形瘤生成能力、分化能力等都与胚胎干细胞极为相似；命名为诱导多能干细胞（Induced Pluripotent Stem Cells，iPS), 这一过程为从体细胞到干细胞的重编程。 四、干细胞移植的发展状况 Neuronal Structure cells immunochemistry markerof dopaminergic neuron Marker of astrocyte Marker of Ectoderm and neuron GFAP: Glial fibrillary acidic protein MAP2: microtubule-associated protein 2 Direct differention into Cardiac Cells Phase-contrast image of iPS cells differentiated into cardiomyocytes. 6.Teratoma Formation from Human iPS Cells three germ layer cells （HE staining） The established human iPS cells are similar to hES cells in many aspects, including morphology, proliferation, feeder dependence, surface markers, gene expression, promoter activities, telomerase activities, in vitro differentiation, and teratoma formation. Any help for your experimental design ? Application of iPSCs produce patient- and disease specific pluripotent stem cell lines disease modeling drug discovery cell replacement strategies Challenges of iPSCs low reprogramming efficiency and rate the reprogramming mechanism is obscure genomic(factors and vectors) integration---chromatin modification---potential tumor formation the evaluation of clinical safty and reliability 体细胞 c-Myc Oct3/4 Klf4 Sox2 Day 1:细胞准备 When human adult fibroblast (HAF) cells have reached 80% confluence, aspirate medium, wash once with PBS, cover cells with 0.05% trypsin, incubate for 5 min at 37 °C. Inactivate trypsin with HAF medium, collect cells in a 50-ml conical tube. Centrifuge the cells at 200g at room temperature for 4 min and discard the supernatant. 从体细胞到干细胞的重编程 Day 1:细胞准备 Resuspend the cel