Primordial germ cell-like cells differentiated in vitro from skin-derived stem cells.
BACKGROUND
We have previously demonstrated that stem cells isolated from fetal porcine skin have the potential to form oocyte-like cells (OLCs) in vitro. However, primordial germ cells (PGCs), which must also be specified during the stem cell differentiation to give rise to these putative oocytes at more advanced stages of culture, were not systematically characterized. The current study tested the hypothesis that a morphologically distinct population of cells derived from skin stem cells prior to OLC formation corresponds to putative PGCs, which differentiate further into more mature gametes.


RESULTS
When induced to differentiate in an appropriate microenvironment, a subpopulation of morphologically distinct cells, some of which are alkaline phosphatase (AP)-positive, also express Oct4, Fragilis, Stella, Dazl, and Vasa, which are markers indicative of germ cell formation. A known differentially methylated region (DMR) within the H19 gene locus, which is demethylated in oocytes after establishment of the maternal imprint, is hypomethylated in PGC-like cells compared to undifferentiated skin-derived stem cells, suggesting that the putative germ cell population undergoes imprint erasure. Additional evidence supporting the germ cell identity of in vitro-generated PGC-like cells is that, when labeled with a Dazl-GFP reporter, these cells further differentiate into GFP-positive OLCs.


CONCLUSIONS
The ability to generate germ cell precursors from somatic stem cells may provide an in vitro model to study some of the unanswered questions surrounding early germ cell formation.
Linher
Katja
K
Department of Animal and Poultry Science, University of Guelph, Guelph, Ontario, Canada.
Dyce
Paul
P
Li
Julang
J
D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D048430:Cell Shape; D019175:DNA Methylation / Q000235:genetics; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D056426:Genetic Loci / Q000235:genetics; D005854:Germ Cells / Q000166:cytology* / Q000378:metabolism; D049452:Green Fluorescent Proteins; D016086:Lentivirus / Q000235:genetics; D009865:Oocytes / Q000166:cytology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012867:Skin / Q000166:cytology*; D013234:Stem Cells / Q000166:cytology*; D034421:Sus scrofa; D014161:Transduction, Genetic
D015415:Biomarkers; D012333:RNA, Messenger; D049452:Green Fluorescent Proteins