Efficient generation of neural-like cells from porcine ovarian putative stem cells – morphological characterization and evaluation of their electrophysiological properties

Wartalski, Kamil; Gorczyca, Gabriela; Wiater, Jerzy; Tabarowski, Zbigniew; Palus-Chramiec, Katarzyna; Setkowicz, Zuzanna; Duda, Małgorzata

doi:10.1016/j.theriogenology.2020.05.034

Cited by 4 publications

(12 citation statements)

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“…Among various animal experimental models, pigs share many similarities with humans in the form of organ size, physiology, and functioning [ 31 ]. The limited ethical dilemmas and importantly, successful isolation of putative stem cells from the ovarian cortex (poPSCs) [ 32 , 33 , 34 ], make pigs the valuable experimental model for not only preclinical assessment for stem cell therapy but also for the purposes of somatic cell nuclear transfer (SCNT)-based cloning in mammals. The efforts undertaken to use poPSCs, which have been subjected to extrinsic anabolic steroid-dependent oncogenic transformation (carcinogenesis), as a completely new source of nuclear donor cells (NDCs) for future research focused on SCNT, have not yet been accomplished.…”

Section: Introductionmentioning

confidence: 99%

Anabolic Steroids-Driven Regulation of Porcine Ovarian Putative Stem Cells Favors the Onset of Their Neoplastic Transformation

Gorczyca

Wartalski

Wiater

et al. 2021

IJMS

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Nandrolone (Ndn) and boldenone (Bdn), the synthetic testosterone analogues with strong anabolic effects, despite being recognized as potentially carcinogenic compounds, are commonly abused by athletes and bodybuilders, which includes women, worldwide. This study tested the hypothesis that different doses of Ndn and Bdn can initiate neoplastic transformation of porcine ovarian putative stem cells (poPSCs). Immunomagnetically isolated poPSCs were expanded ex vivo in the presence of Ndn or Bdn, for 7 and 14 days. Results show that pharmacological doses of both Ndn and Bdn, already after 7 days of poPSCs culture, caused a significant increase of selected, stemness-related markers of cancer cells: CD44 and CD133. Notably, Ndn also negatively affected poPSCs growth not only by suppressing their proliferation and mitochondrial respiration but also by inducing apoptosis. This observation shows, for the first time, that chronic exposure to Ndn or Bdn represents a precondition that might enhance risk of poPSCs neoplastic transformation. These studies carried out to accomplish detailed molecular characterization of the ex vivo expanded poPSCs and their potentially cancerous derivatives (PCDs) might be helpful to determine their suitability as nuclear donor cells (NDCs) for further investigations focused on cloning by somatic cell nuclear transfer (SCNT). Such investigations might also be indispensable to estimate the capabilities of nuclear genomes inherited from poPSCs and their PCDs to be epigenetically reprogrammed (dedifferentiated) in cloned pig embryos generated by SCNT. This might open up new possibilities for biomedical research aimed at more comprehensively recognizing genetic and epigenetic mechanisms underlying not only tumorigenesis but also reversal/retardation of pro-tumorigenic intracellular events.

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Section: Introductionmentioning

confidence: 99%

Anabolic Steroids-Driven Regulation of Porcine Ovarian Putative Stem Cells Favors the Onset of Their Neoplastic Transformation

Gorczyca

Wartalski

Wiater

et al. 2021

IJMS

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“…Successful isolation and in vitro maintenance of ASCs is of increasing importance in the field of applied biology. Our team's previous results (Wartalski et al 2016(Wartalski et al , 2020 Fig. 1 Expression of multipotent/mesenchymal stem cell markers after the third passage, as analyzed by immunofluorescence, Western blotting, and qRT-PCR.…”

Section: Discussionmentioning

confidence: 98%

“…Total protein extraction and Western blot analysis were conducted as previously described (Wartalski et al 2020).…”

Section: Western Blot Analysismentioning

confidence: 99%

“…Several studies conducted in recent years, also by our team, have provided evidence for the presence of ASCs in mammalian ovaries, due to their high diversity also referred to as putative stem cells (PSCs) (Wartalski et al 2016 ; Yazdekhasti et al 2017 ; Patel et al 2018 ). Our latest results (Wartalski et al 2020 ) demonstrated that porcine ovarian putative stem cells (poPSCs) isolated from the ovarian cortex, despite expression of pluripotency markers, are not pluripotent. As poPSCs can differentiate into functional neural-like cells, they may be multipotent, and poPSCs likely represent mesenchymal stem cell (MSC) populations within the ovary.…”

Section: Introductionmentioning

confidence: 99%

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Porcine ovarian cortex-derived putative stem cells can differentiate into endothelial cells in vitro

Wartalski

Gorczyca

Wiater

et al. 2021

Histochem Cell Biol

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Endothelial cells (ECs), the primary component of the vasculature, play a crucial role in neovascularization. However, the number of endogenous ECs is inadequate for both experimental purposes and clinical applications. Porcine ovarian putative stem cells (poPSCs), although not pluripotent, are characterized by great plasticity. Therefore, this study aimed to investigate whether poPSCs have the potential to differentiate into cells of endothelial lineage. poPSCs were immunomagnetically isolated from postnatal pig ovaries based on the presence of SSEA-4 protein. Expression of mesenchymal stem cells (MSCs) markers after pre-culture, both at the level of mRNA: ITGB1, THY, and ENG and corresponding protein: CD29, CD90, and CD105 were significantly higher compared to the control ovarian cortex cells. To differentiate poPSCs into ECs, inducing medium containing vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), insulin-like growth factor (IGF), epidermal growth factor (EGF), ascorbic acid, and heparin was applied. After 14 days, poPSC differentiation into ECs was confirmed by immunofluorescence staining for vascular endothelial cadherin (VECad) and vascular endothelial growth factor receptor-2 (VEGFR-2). Semi-quantitative WB analysis of these proteins confirmed their high abundance. Additionally, qRT-PCR showed that mRNA expression of corresponding marker genes: CDH5, KDR was significantly higher compared with undifferentiated poPSCs. Finally, EC functional status was confirmed by the migration test that revealed that they were capable of positive chemotaxis, while tube formation assay demonstrated their ability to develop capillary networks. In conclusion, our results provided evidence that poPSCs may constitute the MSC population in the ovary and confirmed that they might be a potential source of ECs for tissue engineering.

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“…In this regard, cells derived from various porcine tissues have been viewed as particularly appropriate given the overall similarity in size and physiology of porcine organs compared to those from humans. Indeed, in recent publications, Wartalski and colleagues have shown that porcine ovarian putative stem cells (poPSCs) derived from the ovary cortex from sexually immature pigs display multipotency (Wartalski et al 2016), with the ability under precisely controlled culture conditions to differentiate into neural-like cells (Wartalski et al 2020). Given the need for a source of endothelial cells for tissue engineering and human regenerative medicine, the porcine model, which as mentioned above has been used previously for many xenogeneic procedures (see also for instance Gasek et al 2021), appeared to represent a potential source for these cells.…”

Section: Differentiation Of Porcine Ovarian Mesenchymal Stem Cells Into Endothelial Cellsmentioning

confidence: 99%

In focus in HCB

Taatjes

Roth

2021

Histochem Cell Biol

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Efficient generation of neural-like cells from porcine ovarian putative stem cells – morphological characterization and evaluation of their electrophysiological properties

Cited by 4 publications

References 61 publications

Anabolic Steroids-Driven Regulation of Porcine Ovarian Putative Stem Cells Favors the Onset of Their Neoplastic Transformation

Anabolic Steroids-Driven Regulation of Porcine Ovarian Putative Stem Cells Favors the Onset of Their Neoplastic Transformation

Porcine ovarian cortex-derived putative stem cells can differentiate into endothelial cells in vitro

In focus in HCB

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