GATA transcription in a small rhodamine 123(low)CD34(+) subpopulation of a peripheral blood-derived CD34(-)CD105(+) mesenchymal cell line.
OBJECTIVE
Based on previous animal experiments that suggest the plasticity of peripheral blood-derived, CD34(-) stem cell lines, the aim of this study was to isolate CD34(-) stem cell lines from human peripheral blood cells and obtain evidence of their multipotency and plasticity.


METHODS
Adherent growing cells were isolated from peripheral blood mononuclear cells from a healthy volunteer donor and different cell clones were established after SV40 large-T-antigen-mediated immortalization. The immunophenotype of the cell lines was investigated by flow cytometry. One particular cell clone, V54/2, was stained with rhodamine 123, and the Rh123(low) and Rh123(high) subpopulations were sorted for a reverse transcriptase polymerase chain reaction gene expression survey and distinct differences in morphology and biologic behavior.


RESULTS
The peripheral blood-derived and fibroblast-like cell line V54/2 expressed high levels of CD10 and CD105 and showed only a very low level expression of CD34 (<1.0%) and CD117 (c-kit). Among the entire CD34(-)CD105(+) cell population that transcribed factors such as Myb, Tie-1, and VEGF, there was a small Rh123(low)CD34(+) subpopulation that transcribed significant levels of several members of the GATA family of transcription factors. The morphology of the Rh123(low)CD34(+) (also expressing the P-glycoprotein) was different compared to the Rh123(high)CD34(-) population. Mesenchymal differentiation into glial fibrillary acidic protein (GFAP)(+) glial cells could be shown from the entire CD34(-)CD105(+) cell population.


CONCLUSIONS
The findings provide evidence that it is possible to isolate CD34(-)CD105(+) mesenchymal stem cell lines from human peripheral blood cells that contain a small subpopulation of CD34(+) and GATA-transcribing cells. Those cells are potential hematopoietic progenitors and can be recruited from the CD34(-) stem cell pool. The plasticity of stem cells seems to require essential molecular tools, such as a panel of transcription factors, to respond to the environmental demand within a biologic system.
Conrad
Claudius
C
Institute of Pathology, University of Munich, Germany.
Gottgens
Berthold
B
Kinston
Sarah
S
Ellwart
Joachim
J
Huss
Ralf
R
D000328:Adult; D015703:Antigens, CD; D018952:Antigens, CD34 / Q000032:analysis; D000952:Antigens, Polyomavirus Transforming / Q000502:physiology; D001773:Blood Cells / Q000166:cytology* / Q000378:metabolism; D002454:Cell Differentiation; D002461:Cell Line, Transformed / Q000378:metabolism; D019070:Cell Lineage; D002999:Clone Cells / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics; D000071063:Endoglin; D050981:Erythroid-Specific DNA-Binding Factors; D005456:Fluorescent Dyes / Q000032:analysis; D050989:GATA2 Transcription Factor; D050990:GATA3 Transcription Factor; D050993:GATA6 Transcription Factor; D020869:Gene Expression Profiling; D005914:Globins / Q000096:biosynthesis / Q000235:genetics; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology; D019650:Hematopoietic Stem Cell Mobilization; D006412:Hematopoietic Stem Cells / Q000166:cytology; D006801:Humans; D016130:Immunophenotyping; D008648:Mesoderm / Q000166:cytology* / Q000378:metabolism; D005810:Multigene Family; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000096:biosynthesis / Q000235:genetics; D011956:Receptors, Cell Surface; D020133:Reverse Transcriptase Polymerase Chain Reaction; D020112:Rhodamine 123 / Q000032:analysis; D015534:Trans-Activators / Q000096:biosynthesis / Q000235:genetics; D014157:Transcription Factors / Q000096:biosynthesis* / Q000235:genetics; D019010:Vascular Cell Adhesion Molecule-1 / Q000032:analysis
D015703:Antigens, CD; D018952:Antigens, CD34; D000952:Antigens, Polyomavirus Transforming; D004268:DNA-Binding Proteins; C064206:ENG protein, human; D000071063:Endoglin; D050981:Erythroid-Specific DNA-Binding Factors; D005456:Fluorescent Dyes; D050989:GATA2 Transcription Factor; C494711:GATA2 protein, human; D050990:GATA3 Transcription Factor; C494749:GATA3 protein, human; D050993:GATA6 Transcription Factor; C494759:GATA6 protein, human; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D011956:Receptors, Cell Surface; D015534:Trans-Activators; D014157:Transcription Factors; D019010:Vascular Cell Adhesion Molecule-1; D016179:Granulocyte Colony-Stimulating Factor; D020112:Rhodamine 123; D005914:Globins