Journal de Fertilization: In Vitro - IVF-Worldwide, Médecine de la Reproduction, Génétique & Biologie des Cellules Souches
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Magnetic Isolation and Characterization of Porcine Ovarian Putative Stem Cells (Pscs): An In Vitro Study

Malgorzata Duda

According to the ruling hypothesis the potential source of tumorigenesis might be cancer stem cells (CSCs) resulting from the accumulation of mutations in other types of stem cells (SCs). Therefore we decided to identify and characterize the somatic stem cells commonly called Putative Stem Cells (PSCs) in the porcine postnatal ovary. To isolate ovarian PSCs, an antibody against SSEA-4 conjugated to magnetic beads was used. PSCs were cultured for 7 days. During the culture spontaneously arising embryoid bodies have been observed. Such behaviour under culture condition is characteristic for stem cells. Using immunofluorescence we showed cytoplasmic localization of the Oct 3/4 and Sox-2, pluripotency markers, proteins responsible for maintaining cells in an undifferentiated state. Using immunocytochemistry we showed transmembrane and cytoplasmic localization of the c-Kit receptor and GDF-9 - protein associated with the initiation of cell differentiation in the process of folliculogenesis. We demonstrated also a cytoplasmic and perinuclear location of the Ki-67 protein associated with cell division. PCR method revealed the presence of stem cell markers such as c-Kit, GDF-9 and proliferation marker Ki-67 at the level of the transcript in the putative stem cells (PSCs). Moreover, by the Western blot analysis we also confirmed the presence of markers including c-Kit, GDF-9, Oct-3/4, Sox-2 and Ki-67 in these PSC cells. The results obtained allow to presume that stem cells are actually present in the porcine ovary.