全能干细胞研究及在再生医学中的应用

Nature Reviews Genetics 7, 261-276 (April 2006)
多能干细胞 ：具有无限的自我更新能力和分化成体 内任何种类细胞，包括生殖细胞的潜能。
Embryonic Stem Cells 源自文库胎干细胞 Induced Pluripotent Stem Cells 诱导多能干细胞
Directed Differentiation Transplantation
Concepts
Stem Cells Pluripotent Stem Cells
Research on Pluripotent Stem Cells
Derivation and Culture Characterization
Sun, et al.
Sun, et al.
The first human ESC line was established in 1998 by James A. Thomson.
Science 282: 1145, 1998
hESC maintenance (Feeder+)
Feeder cells producing Activin A， Adding bFGF
全能干细胞研究及 在再生医学中的应用
Concepts
Stem Cells Pluripotent Stem Cells
Research on Pluripotent Stem Cells
Derivation and Culture Characterization
Application of Pluripotent Stem Cells
Blastocyst Fetus
Fetus
Totipotent
Reprogramming
Pluripotent
Multi-potent or Uni-potent
Methylation X-inactivation
Imprint status Marker expression
Regulatory gene expression Position Information
Basal Culture Medium/ Human Serum/ Human Gelatin/ Human Trypsin
PLoS ONE August 2009 | Volume 4 | Issue 8 | e6724
SHhES6
SHhES7
Image by Catherine Twomey Understanding Stem Cells: An Overview of the Science and Issues
Symmetric division
Asymmetric division
Developmental potential Self-renewal capacity
Li et al. In Vitro Cell.Dev.Biol.-Animal (2010) 46:186–191
Feeder-free culture hES cells
Feeder cells derived from human tissue:
Fetal muscle, fetal skin Adult fallopian tubule epithelial cells Foreskin fibroblast Adult marrow cells Adult lung Uterine endometrium hES cell derived fibroblast
Directed Differentiation Transplantation
Embryonic Stem Cells
来源于哺乳类动物着床前胚胎内细胞团的细胞
Sir Martin John Evans
The first to culture mouse ESCs in 1981 2007 Nobel Laureate
Application of Pluripotent Stem Cells
Directed Differentiation Transplantation
Reproduce Itself Generation one or more differentiated descendant cell types
LIF + BMP
LIF- and BMP-free (2i)
The fibroblast growth factor/extracellular signalrelated kinase (Fgf/Erk1/2) inhibitor---inhibit differentiation Glycogen synthase kinase 3 (GSK3) inhibitor --increase b-catenin levels
Feeder Cells + Serum
Feeder Cells provide leukemia inhibitory factor(LIF) Serum provides bone morphogenetic proteins (BMPs)
Feeder Cells- and Serum-free
hESC maintenance (Feeder-)
Matrigel® (hESC Qualified Matrix) mTeSRTM1 or TeSRTM-E8TM
hESC Passage
Mechanical split or enzymes Combination of enzyme and manual splitting Enzyme with Rock inhibitor for single cell passage
Concepts
Stem Cells Pluripotent Stem Cells
Research on Pluripotent Stem Cells
Derivation and Culture Characterization
Application of Pluripotent Stem Cells