Using human stem cells to understand & treat disease
PLURIPOTENT HUMAN CELLS
HUMAN GENETICS
GLIOBLASTOMA Josh Chenoweth, Paul Tesar & NIH stem cell facility
TERATOCARCINOMA
LIVER DEVELOPMENT & Jong-Hoon Kim (NIH & Korea University) FUNCTION Controlling embryonic stem cell differentiation in the mouse
A Mdm2 DAPI E3.5
Tr-ectoderm
ICM
B Epiblast E5.5 MEF mES mES mEpiSmEpiS mES
AlkIn AlkIn AlkIn JakIn
mES mEpiS hES
Mdm2 Trophectoderm
C E5.5 E5.5 epiblast embryo + 3 days in vitro
Epiblast
Mdm2 expression in the E3.5 (A) and E5.5 embryo (B). The E5.5 embryo prior to dissection of the epiblast Tesar et al., Nature, 2007 and the epiblast after 3 days in culture (C). Cell types in the early embryo Controlling human embryonic stem cells
Days Whole transcriptome characterization of undifferentiated hES cells after Zygote fertiliz ation
E2.5 Morula Mouse ES
E3.5 Inner Cell Mass
Trophectoderm
Human ES E4.5 Other labs Implant Epiblast Extra-emb. NIH E5.5 Endoderm
E6.5 Mes-endoderm Embryoid Body
Ectoderm Mesoderm Endoderm Controlling differentiation of human embryonic stem cells
Whole transcriptome characterization of hES cells
Human ES
Adult human tissue
Embryoid Bodies Controlling differentiation of human iPS cells
Whole transcriptome characterization of hES cells
ES iPS Parental cells Fibroblasts
iPS EBs
Adult human tissue
Embryoid Bodies Polymorphic levels of gene expression – Lung & prostate cancer
Glutathione S transferase (GSTT1)
ES iPS
BG03
ES06
GSTT1 Expression UC01 GSTT1 Copy Number
Agilent: CGH Analytics used to assess copy number Polymorphic levels of gene expression – Glioblastoma & Teratocarcinoma
MGMT +ve endothelial cells
–ve
Expression melanoma brain metastasis MGMT
MGMT
–ve & +ve breast cancer Expression MGMT +++ve lung cancer
Ingold B, Schraml P, Heppner FL Moch H, MGMT Promoter Methylation Homogeneous MGMT immunoreactivity correlates with unmethylated MGMT promoter status in brain metstases of various solid tumors PLoS One 2009 Generating Functional Hepatocytes
Human Hepatocytes HumanAlbumin
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