Medium for human embryonic stem cells [课件].pptVIP

Culture of Human Embryonic Stem Cells (hESC)人胚胎干细胞培养 What are Stem Cells? Embryonic stem cells are immortal stem cells lines of embryonic origin, which are isolated from the inner cell mass of the human blastocyst. Thse unique cells are characterized by their ability to be continuously cultured in an undifferentiated state, without losing their development potential to differentiate into each cell in the adult body 胚胎干细胞是在人胚胎发育早期——囊胚（受精后约5—7天）中未分化的细胞。 囊胚含有约140个细胞，外表是一层扁平细胞，称滋养层，可发育成胚胎的支持组织 如胎盘等。中心的腔称囊胚腔，腔内一侧的细胞群，称内细胞群，这些未分化的细胞 可进一步分裂、分化，发育成个体。内细胞群在形成内、中、外三个胚层时开始分化 。每个胚层将分别分化形成人体的各种组织和器官。如外胚层将分化为皮肤、眼睛和 神经系统等，中胚层将形成骨骼、血液和肌肉等组织，内胚层将分化为肝、肺和肠等 。由于内细胞群可以发育成完整的个体，因而这些细胞被认为具有全能性。当内细胞 群在培养皿中培养时，我们称之为胚胎干细胞。  Embryonic Stem (ES) Cells 胚胎干细胞 Derived from early embryos (Inner Cell Mass 内细胞群 ) Advantages 优点 Easy to isolate 容易分离出来 Show full potential for all cell types 可分化成所有种类细胞 Extended cell growth capabilities 扩增的细胞生长能力 Disadvantages 缺点 Significant ethical issues 显著的伦理道德问题 Adult Stem Cells成人干细胞 derived from adult tissue 来自成人组织 (hematopoietic 造血组织, mesenchymal 间页细胞组织) Includes umbilical cord stem cells 包括脐带干细胞 Advantages 优点 - can be isolated from a patient, expanded in culture and given back to the same patient (no rejection issues) 可以取自患者，培养扩增后返回体内（无排斥） Disadvantages 缺点 - very rare in the body 体内非常少 - hard to identify 难于辨识 - limited developmental potential 有限的扩展潜力 - very limited growth potential 非常有限的生长潜力 The Promise of Stem Cell 干细胞的意义 Culture of hESC’s 人干细胞培养 Serum-containing medium: 含血清培养基 Dulbecco’s Modified Eagle’s Medium (DMEM) 20% Foetal Bovine Serum 2mM L-glutamine Non-Essential Amino Acids Β-Mercaptoethanol 巯基乙醇 Culture of hESC’s 人胚胎干细胞培养 Feeder layer 滋养层 Mouse Embryo Fibroblasts (MEF) 鼠胚胎纤维原细胞 Human Foreskin Fibroblasts (HFF) 人包皮纤维原细胞 Feeder layer free 无滋养层 MEF/HFF conditioned medium MEF/HFF处理过培养基 Human serum matrix 人血清基质 Matrigel 基质胶 HFF matrix HFF 基质 Development of NutriStem hESC medium for Human Embryonic Stem Cells NutriStem hESCACF SFM