Isolation of Small SSEA-4-Positive Putative Stem Cells from the Ovarian Surface Epithelium of Adult Human Ovaries by Two Different Methods

Virant‐Klun, Irma; Skutella, Thomas; Hren, Matjaž; Gruden, Kristina; Cvjetičanin, Branko; Vogler, Andrej; Šinkovec, Jasna

doi:10.1155/2013/690415

Cited by 65 publications

(69 citation statements)

References 36 publications

(37 reference statements)

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“…It is difficult to isolate and grow a large population of cells expressing the markers tested. According to literature reports, human cells of the ovarian surface epithelium (OSE) with an epitope of SSEA-4 demonstrate the expression of stem cell markers such as Oct-3/4, Nanog and Sox-2 [14]. Herein our experiments confirmed that PSCs can be isolated by magnetic beads due to the presence of an epitope SSEA-4 also from the porcine ovary.…”

Section: Discussionsupporting

confidence: 82%

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Magnetic Isolation and Characterization of Porcine Ovarian Putative Stem Cells (Pscs): An In Vitro Study

Duda¹

2016

JFIV Reprod Med Genet

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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.

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Section: Discussionsupporting

confidence: 82%

“…In normal development, SSEA4 is expressed by the majority of pregonadal primordial germ cells (PGCs) and is progressively downregulated when they enter into meiosis in the embryonic ovary [13]. However, it has been stated that the human SSEA4-positive PSCs from ovarian cell cultures are related to ESCs and cells of the germinal lineage [14].…”

Section: Discussionmentioning

confidence: 99%

Magnetic Isolation and Characterization of Porcine Ovarian Putative Stem Cells (Pscs): An In Vitro Study

Duda¹

2016

JFIV Reprod Med Genet

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“…Similarly, Virant-Klun's et al (2013) has reported that OSE harbors stem cells, which spontaneously differentiate into oocyte-like structures during in vitro culture. Our group also observed spontaneous differentiation of OSE cells into oocyte-like structures and that the epithelial cells undergo epithelial-mesenchymal transition to form granulosa-like cells that surround the developing germ cells resulting in follicle assembly (Parte et al 2011).…”

Section: R41mentioning

confidence: 89%

FSH–FSHR3–stem cells in ovary surface epithelium: basis for adult ovarian biology, failure, aging, and cancer

Bhartiya

Singh

2015

Reproduction

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Despite extensive research, genetic basis of premature ovarian failure (POF) and ovarian cancer still remains elusive. It is indeed paradoxical that scientists searched for mutations in FSH receptor (FSHR) expressed on granulosa cells, whereas more than 90% of cancers arise in ovary surface epithelium (OSE). Two distinct populations of stem cells including very small embryonic-like stem cells (VSELs) and ovarian stem cells (OSCs) exist in OSE, are responsible for neo-oogenesis and primordial follicle assembly in adult life, and are modulated by FSH via its alternatively spliced receptor variant FSHR3 (growth factor type 1 receptor acting via calcium signaling and the ERK/MAPK pathway). Any defect in FSH-FSHR3-stem cell interaction in OSE may affect folliculogenesis and thus result in POF. Ovarian aging is associated with a compromised microenvironment that does not support stem cell differentiation into oocytes and further folliculogenesis. FSH exerts a mitogenic effect on OSE and elevated FSH levels associated with advanced age may provide a continuous trigger for stem cells to proliferate resulting in cancer, thus supporting gonadotropin theory for ovarian cancer. Present review is an attempt to put adult ovarian biology, POF, aging, and cancer in the perspective of FSH-FSHR3-stem cell network that functions in OSE. This hypothesis is further supported by the recent understanding that: i) cancer is a stem cell disease and OSE is the niche for ovarian cancer stem cells; ii) ovarian OCT4-positive stem cells are regulated by FSH; and iii) OCT4 along with LIN28 and BMP4 are highly expressed in ovarian cancers.Reproduction (2015) 149 R35-R48

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“…To elucidate whether the oocytes generated were truly derived from mitotically active PSCs, and did not instead represent oocytes derived from primary ovarian cells, we isolated and purified PSCs by SSEA4-based magnetic bead sorting, as small SSEA4-positive cells from human ovarian cell cultures are reportedly related to ESCs and cells of the germinal lineage (Virant-Klun et al, 2013), and small porcine PSCs showed cytoplasmic expression of SSEA4 (supplementary material Fig. S4A).…”

Section: Arrows) (C) After 2 Weeks Thementioning

confidence: 99%

Identification and characterization of putative stem cells in the adult pig ovary

et al. 2014

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Recently, the concept of 'neo-oogenesis' has received increasing attention, since it was shown that adult mammals have a renewable source of eggs. The purpose of this study was to elucidate the origin of these eggs and to confirm whether neo-oogenesis continues throughout life in the ovaries of the adult mammal. Adult female pigs were utilized to isolate, identify and characterize, including their proliferation and differentiation capabilities, putative stem cells (PSCs) from the ovary. PSCs were found to comprise a heterogeneous population based on c-kit expression and cell size, and also express stem and germ cell markers. Analysis of PSC molecular progression during establishment showed that these cells undergo cytoplasmic-to-nuclear translocation of Oct4 in a manner reminiscent of gonadal primordial germ cells (PGCs). Hence, cells with the characteristics of early PGCs are present or are generated in the adult pig ovary. Furthermore, the in vitro establishment of porcine PSCs required the presence of ovarian cell-derived extracellular regulatory factors, which are also likely to direct stem cell niche interactions in vivo. In conclusion, the present work supports a crucial role for c-kit and kit ligand/stem cell factor in stimulating the growth, proliferation and nuclear reprogramming of porcine PSCs, and further suggests that porcine PSCs might be the culture equivalent of early PGCs.

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Isolation of Small SSEA-4-Positive Putative Stem Cells from the Ovarian Surface Epithelium of Adult Human Ovaries by Two Different Methods

Cited by 65 publications

References 36 publications

Magnetic Isolation and Characterization of Porcine Ovarian Putative Stem Cells (Pscs): An In Vitro Study

Magnetic Isolation and Characterization of Porcine Ovarian Putative Stem Cells (Pscs): An In Vitro Study

FSH–FSHR3–stem cells in ovary surface epithelium: basis for adult ovarian biology, failure, aging, and cancer

Identification and characterization of putative stem cells in the adult pig ovary

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