Magnetic Isolation and Characterization of Porcine Ovarian Putative Stem Cells (Pscs): An In Vitro Study
- *Corresponding Author:
- Malgorzata Duda
Department of Endocrinology
Institute of Zoology, Jagiellonian University
Gronostajowa 9, 30-387 Krakow, Poland
Tel: +48 12 664 50 32
Fax: +48 126-645-098
E-mail: [email protected]
Received date July 14, 2016; Accepted date August 25, 2016; Published date September 02, 2016
Citation: Wartalski K, Tabarowski Z, Duda M (2016) Magnetic Isolation and Characterization of Porcine Ovarian Putative Stem Cells (Pscs): An In Vitro Study.J Fertil In Vitro IVF Worldw Reprod Med Genet Stem Cell Biol 4:191. doi:10.4172/2375-4508.1000191
Copyright: © 2016 Wartalski K, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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.