Differential responses to retinoic acid and endocrine disruptor compounds of subpopulations within human embryonic stem cell lines.
The heterogeneous nature of stem cells is an important issue in both research and therapeutic use in terms of directing cell lineage differentiation pathways, as well as self-renewal properties. Using flow cytometry we have identified two distinct subpopulations by size, large and small, within cultures of human embryonic stem (hES) cell lines. These two cell populations respond differentially to retinoic acid (RA) differentiation and several endocrine disruptor compounds (EDC). The large cell population responds to retinoic acid differentiation with greater than a 50% reduction in cell number and loss of Oct-4 expression, whereas the number of the small cell population does not change and Oct-4 protein expression is maintained. In addition, four estrogenic compounds altered SSEA-3 expression differentially between the two cell subpopulations changing their ratios relative to each other. Both populations express stem cell markers Oct-4, Nanog, Tra-1-60, Tra-1-80 and SSEA-4, but express low levels of differentiation markers common to the three germ layers. Cloning studies indicate that both populations can revive the parental population. Furthermore, whole genome microarray identified approximately 400 genes with significantly different expression between the two populations (p<0.01). We propose the differential response to RA in these populations is due to differential gene expression of Notch signaling members, CoupTF1 and CoupTF2, chromatin remodeling and histone modifying genes that render the small population resistant to RA differentiation. The findings that hES cells exist as heterogeneous populations with distinct responses to differentiation signals and environmental stimuli will be relevant for their use for drug discovery and disease therapy.
Chromatin and Gene Expression Section, Laboratory of Molecular Carcinogenesis, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC 27709, USA.
Lois A
LA
Annab
Carl D
CD
Bortner
Marie I
MI
Sifre
Jennifer M
JM
Collins
Ruchir R
RR
Shah
Darlene
D
Dixon
H
H
Karimi Kinyamu
Trevor K
TK
Archer
D000943:Antigens, Differentiation / Q000378:metabolism; D000954:Antigens, Surface / Q000096:biosynthesis; D015295:Antigens, Tumor-Associated, Carbohydrate / Q000096:biosynthesis; D047310:Apigenin / Q000494:pharmacology; D051837:COUP Transcription Factor II / Q000096:biosynthesis; D002453:Cell Cycle / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D007631:Chlordecone / Q000494:pharmacology; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D052244:Endocrine Disruptors / Q000494:pharmacology*; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D018398:Homeodomain Proteins / Q000096:biosynthesis; D006801:Humans; D044949:Kaempferols / Q000494:pharmacology; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3 / Q000096:biosynthesis; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D011509:Proteoglycans / Q000096:biosynthesis; D015398:Signal Transduction / Q000187:drug effects; D055746:Stage-Specific Embryonic Antigens / Q000096:biosynthesis; D013629:Tamoxifen / Q000494:pharmacology; D014212:Tretinoin / Q000494:pharmacology*
D000943:Antigens, Differentiation; D000954:Antigens, Surface; D015295:Antigens, Tumor-Associated, Carbohydrate; D047310:Apigenin; D051837:COUP Transcription Factor II; D007631:Chlordecone; D052244:Endocrine Disruptors; D018398:Homeodomain Proteins; D044949:Kaempferols; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3; D011509:Proteoglycans; D055746:Stage-Specific Embryonic Antigens; D013629:Tamoxifen; D014212:Tretinoin; C486780:NANOG protein, human; C107064:TRA-1-60 antigen, human; C052924:stage-specific embryonic antigen-3; C066584:stage-specific embryonic antigen-4; C006552:kaempferol; C095569:NR2F2 protein, human