Derivation and Characterization of Rat Embryonic Stem Cells-复旦《医用分子遗传学》课件.ppt

Derivation and Characterization of Rat Embryonic Stem Cells Characteristics of Embryonic Stem Cells Establishment of pluripotent embryonic cell (like) lines Mouse＊, Human Monkey, Rat, Pig, Sheep… Differences Between Human and Mouse ES Cells RAT Model for biomedical research, physiology, toxicology studies… 3I medium Ying et al, Nature ,2008, 453(7194):519-523 Method Fig. 1 Culture of blastocysts in different medium A: LIF+Serum; B: N2B27; C: N2B27+3I Fig. 2 Derivation and maintain of rat ES cells from rat blastocyst Fig. 3 Immunostaining for pluripotency marker in rat ES cells Fig. 5 Immunofluorescence staining for cell surface marker GEO Number: GSE13681 Commonly Used Functional Criteria In vitro differentiation Fig.7 In vitro differentiation of rat ES cells Fig.7 In vitro differentiation of rat ES cells Fig.8 RT-PCR analysis of gene expression in rat ES cells and EBs Fig.8 Immunostaing confirming in vitro differentiation Fig.9 X inactivation occurs during differentiation of famale rat ES cells Production of Chimera Fig.12 Chimeric rat produced by injection of DAc2 into SD blastocysts LIF/STAT3 signaling in rat ES cells Fig. 16 Response of rat ES cells to LIF stimulation  Fig. 17 Positive effect of LIF on rat ES cells  Fig. 21 Bisulfite sequencing of DNA methylation of the Oct4 and Nanog promoter regions 动物实验 分子生物学实验 Thank You! Fig.15 Genotyping analysis of germline rats and its pups A: 153bp; a: 134bp D: SSLP (Simple Sequence Length Polymorphism) -LIF +LIF Fig. 18 Elevation of LIF/STAT3 signaling in rat ES cells  Fig. 19 Rat ES cells grows on L cells  Fig. 20 qRT-PCR analysis expression of Integrins in Rat ES cells C U Fig. 22 Bivalent domains in rat ES cells and rat fibroblast cells Fig. 22 Bivalent domains in rat ES cells and rat fibroblast cells * * * Serum, feeder, CDM, bFGF, Noggin, Activin A LIF, BMP4, Serum, feeder Culture Condition SSEA-3, SSEA-4, TRA-1- 60,GCTM-2, CD9 SSEA-1, CD9 Cell-surface Marker hES mES