Table 1

Comparison of different methods and approaches for the analysis of human ES-cell gene expression


Sperger et al. [1]
Sato et al. [2]
Richards et al. [3]
Abeyta et al. [4]
Zeng et al. [5]

Human ES-cell lines used
H1, H7, H9, H13, H14
H1
HES3, HES4
H9, HSF-1, HSF-6
BG01, BG02
Culture conditions
MEFs
Matrigel
MEFs
MEFs
MEFs
Method of ES-cell isolation
Treatment with collagenase until colonies lifted off the MEFs
Treatment with dispase until cells were free of MEFs
Microdissection to free colonies of MEFs
Mechanical dissection of colonies from MEFs, then collagenase treatment
Trypsinization
Arrays used
Stanford microarrays
Affymetrix arrays (hU133A and mouse U74Av2)
SAGE
Affymetrix arrays (hU133A and hU133B)
Custom 16,659-spot 70-bp oligonucleotide array
Cells compared
hEC, hES and seminoma
hES and published mES [16]
hES and hES and additional SAGE libraries [24]
hES and hES versus published mES [16]
hES and hES versus mES [28]
Primary subtraction method
Somatic and cancer cell lines
Differentiated hES cells
None
None
Pooled human RNA
Software/analysis used
Significance analysis of microarrays (SAM) [25]
dChip and MAS 4.0 (Affymetrix)
Comparison of two SAGE resources with SAGE 2000 [26,27]
MAS 5.0 (Affymetrix)
Gene Pix (Axon Instruments)
Number of genes enriched in human ES cells
1,760
918
8,341
7,385
373
Candidate pluripotency genes*
565
227
192
76
92
Confirmation of gene expression using
RT-PCR
RT-PCR
RT-PCR
Quantitative RT-PCR
RT-PCR

*Candidate pluripotency genes are defined as genes that are found only in all pluripotent cell lines examined in each study. Abbreviations: hEC, human embryonal carcinoma cells; hES, human embryonic stem cells; MEFs, mouse embryonic fibroblasts; mES, mouse embryonic stem cells; RT-PCR, reverse-transcriptase-coupled PCR; SAGE, serial analysis of gene expression.

Pyle et al. Genome Biology 2004 5:235   doi:10.1186/gb-2004-5-8-235