Transcriptional analysis of early lineage commitment in human embryonic stem cells.
BACKGROUND
The mechanisms responsible for the maintenance of pluripotency in human embryonic stem cells, and those that drive their commitment into particular differentiation lineages, are poorly understood. In fact, even our knowledge of the phenotype of hESC is limited, because the immunological and molecular criteria presently used to define this phenotype describe the properties of a heterogeneous population of cells.


RESULTS
We used a novel approach combining immunological and transcriptional analysis (immunotranscriptional profiling) to compare gene expression in hESC populations at very early stages of differentiation. Immunotranscriptional profiling enabled us to identify novel markers of stem cells and their differentiated progeny, as well as novel potential regulators of hESC commitment and differentiation. The data show clearly that genes associated with the pluripotent state are downregulated in a coordinated fashion, and that they are co-expressed with lineage specific transcription factors in a continuum during the early stages of stem cell differentiation.


CONCLUSIONS
These findings, that show that maintenance of pluripotency and lineage commitment are dynamic, interactive processes in hESC cultures, have important practical implications for propagation and directed differentiation of these cells, and for the interpretation of mechanistic studies of hESC renewal and commitment. Since embryonic stem cells at defined stages of commitment can be isolated in large numbers by immunological means, they provide a powerful model for studying molecular genetics of stem cell commitment in the embryo.
Laslett
Andrew L
AL
Monash Institute of Medical Research, Monash University and the Australian Stem Cell Centre, Clayton, Victoria Australia. andrew.laslett@stemcellcentre.edu.au <andrew.laslett@stemcellcentre.edu.au>
Grimmond
Sean
S
Gardiner
Brooke
B
Stamp
Lincon
L
Lin
Adelia
A
Hawes
Susan M
SM
Wormald
Sam
S
Nikolic-Paterson
David
D
Haylock
David
D
Pera
Martin F
MF
D000818:Animals; D018929:Cell Culture Techniques; D002460:Cell Line; D019070:Cell Lineage; D053595:Embryonic Stem Cells / Q000166:cytology*; D019084:Fluorescent Antibody Technique, Indirect; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D005819:Genetic Markers; D006801:Humans; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis; D039904:Pluripotent Stem Cells / Q000166:cytology*; D011994:Recombinant Proteins / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012795:Sialoglycoproteins / Q000235:genetics
D005819:Genetic Markers; D011994:Recombinant Proteins; D012795:Sialoglycoproteins; C001643:podocalyxin