Activation of ectopic Oct-4 and Rex-1 promoters in human amniotic fluid cells.
Recently, amniotic fluid was suggested as a new source for stem-cell research and tissue engineering approaches. In order to enable isolation of stem cells and establishment of lines of such cells with an undifferentiated phenotype we have introduced green fluorescent protein regulated by the promoters of the stem cell-specific genes, Oct-4 or Rex-1, into human amniotic fluid cells. For the introduction of DNA into human amniotic fluid cells, we have optimized a specific transfection protocol. We found that human amniotic fluid contains cell populations which are able to activate these promoters. These undifferentiated cells expressing green fluorescent protein can be analysed on a flow cytometer. In addition, we have introduced a plasmid harboring a neomycin-resistance gene under the control of the Oct-4 promoter. G418 selection allowed the isolation of undifferentiated stem cells expressing Oct-4 protein out of human amniotic fluid samples. Our findings confirm the existence of stem cells within amniotic fluid. In addition, the ability to transfect human amniotic fluid cells and to isolate stem-cell marker-positive cells will provide the means to study and manipulate these cells for the purpose of basic and applied research.
Karlmark
Karlin R
KR
Medical Genetics, Obstetrics and Gynecology, Medical University of Vienna, 1090 Vienna, Austria.
Freilinger
Angelika
A
Marton
Erika
E
Rosner
Margit
M
Lubec
Gert
G
Hengstschläger
Markus
M
D000653:Amniotic Fluid / Q000166:cytology* / Q000378:metabolism*; D002352:Carrier Proteins / Q000235:genetics*; D005260:Female; D017930:Genes, Reporter / Q000235:genetics; D006801:Humans; D051741:Kruppel-Like Transcription Factors; D050814:Octamer Transcription Factor-3 / Q000235:genetics*; D009928:Organ Specificity; D011247:Pregnancy; D011401:Promoter Regions, Genetic / Q000235:genetics*; D013234:Stem Cells / Q000378:metabolism; D015533:Transcriptional Activation / Q000235:genetics*; D014162:Transfection
D002352:Carrier Proteins; D051741:Kruppel-Like Transcription Factors; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; C507976:ZFP42 protein, human