Ectopic overexpression of Nanog induces tumorigenesis in non-tumorous fibroblasts

2016 ◽  
Vol 397 (3) ◽  
pp. 249-255 ◽  
Author(s):  
Yo Seph Park ◽  
Judee Grace E. Nemeño ◽  
Na Young Choi ◽  
Jeong Ik Lee ◽  
Kisung Ko ◽  
...  
Keyword(s):  
Stem Cells ◽  
Morphological Changes ◽  
Embryonic Stem ◽  
Mouse Embryonic Fibroblast ◽  
Tumor Formation ◽  
Proliferation Rate ◽  
Fibroblast Cells ◽  
Reprogramming Factors ◽  
Embryonic Fibroblast

Abstract Key regulatory genes in pluripotent stem cells are of interest not only as reprogramming factors but also as regulators driving tumorigenesis. Nanog is a transcription factor involved in the maintenance of embryonic stem cells and is one of the reprogramming factors along with Oct4, Sox2, and Lin28. Nanog expression has been detected in different types of tumors, and its expression is a poor prognosis for cancer patients. However, there is no clear evidence that Nanog is functionally involved in tumorigenesis. In this study, we induced overexpression of Nanog in mouse embryonic fibroblast cells and subsequently assessed their morphological changes, proliferation rate, and tumor formation ability. We found that Nanog overexpression induced immortalization of mouse embryonic fibroblast cells (MEFs) and increased their proliferation rate in vitro. We also found that formation of tumors after subcutaneous injection of retroviral-Nanog infected MEFs (N-MEFs) into athymic mouse. Cancer-related genes such as Bmi1 were expressed at high levels in N-MEFs. Hence, our results demonstrate that Nanog is able to transform normal somatic cells into tumor cells.

Calcium hypochlorite on mouse embryonic fibroblast cells (NIH3T3) in vitro cytotoxicity and genotoxicity: MTT and comet assay

2020 ◽  
Vol 47 (7) ◽  
pp. 5377-5383
Author(s):  
Şehnaz Yilmaz ◽  
Oguz Yoldas ◽  
Aysin Dumani ◽  
Gizem Guler ◽  
Seda Ilgaz ◽  
...  
Keyword(s):  
Comet Assay ◽  
Mouse Embryonic Fibroblast ◽  
In Vitro Cytotoxicity ◽  
Fibroblast Cells ◽  
Calcium Hypochlorite ◽  
Embryonic Fibroblast

scholarly journals ФЕНОТИПОВІ ТА МОРФОЛОГІЧНІ ЗМІНИ КУЛЬТУРИ КЛІТИН КІСТКОВОГО МОЗКУ ЩУРІВ В ПРОЦЕСІ ЇХ КУЛЬТИВУВАННЯ

2016 ◽  
Vol 18 (2(66)) ◽  
pp. 126-132
Author(s):  
A.I. Mazurkiewicz ◽  
V.V. Kovpak ◽  
O.S. Kovpak
Keyword(s):  
Stem Cells ◽  
Cell Culture ◽  
Bone Marrow ◽  
Bone Marrow Cells ◽  
Morphological Changes ◽  
Embryonic Stem ◽  
In Vitro Cultivation ◽  
Rat Bone ◽  
Marrow Cells

Bone marrow is the only adult tissue which normally consists of immature undifferentiated and low differentiated cells which called stem cells and they are similar in structure to embryonic stem cells. But literature data analysis doesn't give an unambiguous answer regarding phenotypic and morphological changes of bone marrow cells culture of rats during their in vitro cultivation which necessitated further research.Investigate phenotypic and morphological changes of bone marrow cells culture of rats during their in vitro cultivation from first to fourth passage.We were used in these research bone marrow cells of rats from the first to the fourth passages. Microscopic analysis and evaluation morphological changes of bone marrow cells culture of rats during cultivation were carried out using inverted microscope Axiovert 40. Control of changes phenotype was performed by detecting CD markers (CD10, CD38, CD34, CD45, CD48, CD54, CD56, CD66e, CD96, CD227, CD326, pan–keratin). The evaluation was performed by the semi– quantitative method (H–Score).The research of primary culture of rat bone marrow cells showed that it morphologically heterogeneous, noted the small number of cells polygonal shape, surrounded by the fibroblast cells. During the cultivation cell culture becomes more homogenous at the expense of fibroblast–like cells. As a result of occurred the transition process from heterogeneous culture in zero passage to the most homogeneous culture in 4 passage. Immunophenotyping population of cell culture derived from rat bone marrow, revealed a high level of expression of pan–keratin; moderate level – CD34, CD48, CD66e, CD95; low level – CD38, CD45, CD56, CD227, CD326; lack of expression – CD10, CD54. Change of the expression of surface markers varies in each passage CD48, CD66e, CD95 increased significantly; CD38, SD45, SD326, pan–keratin reduced significantly. The markers CD34, CD 56, CD 227 were expressed on the one level from the first to the fourth passage. The expression of the CD10, CD54 markers during the study period was not identified.

scholarly journals In Vitro Evaluation of the Biocompatibility and Stability of Fluorescent Dil Dye on Mouse Embryonic Fibroblast Cells

2019 ◽  
Vol In Press (In Press) ◽  
Author(s):  
Marzyeh Haghshenas ◽  
Elham Hoveizi ◽  
Tayebeh Mohammadi ◽  
Seyed Reza Kazemi Nezhad
Keyword(s):  
Mouse Embryonic Fibroblast ◽  
Fibroblast Cells ◽  
In Vitro Evaluation ◽  
Embryonic Fibroblast

scholarly journals The Effect of Rhodamine-Derived Superparamagnetic Maghemite Nanoparticles on the Motility of Human Mesenchymal Stem Cells and Mouse Embryonic Fibroblast Cells

Molecules ◽  
2019 ◽  
Vol 24 (7) ◽  
pp. 1192
Author(s):  
Larisa Baiazitova ◽  
Josef Skopalik ◽  
Jiri Chmelik ◽  
Inna Zumberg ◽  
Vratislav Cmiel ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Proliferation ◽  
Mesenchymal Stem Cells ◽  
Human Mesenchymal Stem Cells ◽  
Cell Types ◽  
Mouse Embryonic Fibroblast ◽  
Fibroblast Cells ◽  
Maghemite Nanoparticles ◽  
Safety Research ◽  
Embryonic Fibroblast

Nanoparticles have become popular in life sciences in the last few years. They have been produced in many variants and have recently been used in both biological experiments and in clinical applications. Due to concerns over nanomaterial risks, there has been a dramatic increase in investigations focused on safety research. The aim of this paper is to present the advanced testing of rhodamine-derived superparamagnetic maghemite nanoparticles (SAMN-R), which are used for their nontoxicity, biocompatibility, biodegradability, and magnetic properties. Recent results were expanded upon from the basic cytotoxic tests to evaluate cell proliferation and migration potential. Two cell types were used for the cell proliferation and tracking study: mouse embryonic fibroblast cells (3T3) and human mesenchymal stem cells (hMSCs). Advanced microscopic methods allowed for the precise quantification of the function of both cell types. This study has demonstrated that a dose of nanoparticles lower than 20 µg·cm−2 per area of the dish does not negatively affect the cells’ morphology, migration, cytoskeletal function, proliferation, potential for wound healing, and single-cell migration in comparison to standard CellTracker™ Green CMFDA (5-chloromethylfluorescein diacetate). A higher dose of nanoparticles could be a potential risk for cytoskeletal folding and detachment of the cells from the solid extracellular matrix.

scholarly journals Exosomes derived from embryonic stem cells inhibit doxorubicin and inflammation-induced pyroptosis in muscle cells

2018 ◽  
Vol 96 (3) ◽  
pp. 304-307 ◽  
Author(s):  
Zahra Tavakoli Dargani ◽  
Reetu Singla ◽  
Taylor Johnson ◽  
Rakesh Kukreja ◽  
Dinender K. Singla
Keyword(s):  
Inflammatory Cytokine ◽  
In Vitro Model ◽  
Embryonic Stem ◽  
Mouse Embryonic Fibroblast ◽  
Caspase 1 ◽  
Key Players ◽  
Vitro Model ◽  
Embryonic Fibroblast ◽  
Muscle Toxicity

Doxorubicin (Dox) is an effective anticancer drug. Unfortunately, it causes cardiac and muscle toxicity due to increased oxidative stress and inflammation; however, it remains unknown whether Dox induces “pyroptosis” — an inflammation-mediated cell death. We investigated whether Dox induces pyroptosis in mouse soleus muscle (Sol 8) cells in vitro and to show the protective effect of embryonic stem cell exosomes (ES-exos) on pyroptosis. Dox and inflammation-induced in vitro model was generated. Pyroptosis was confirmed using immunohistochemistry (with putative markers caspase-1, IL-1β, and pro-inflammatory cytokine IL-18) and Western blotting of caspase-1 and IL-1β. The results show significant increase in the expression of caspase-1, IL-1β, and IL-18 following treatment with Dox, which was inhibited by ES-exos but not mouse embryonic fibroblast exosomes. Moreover, GW4869 compound inhibited functional activity of ES-exos, suggesting these vesicles are key players in the inhibition of pyroptosis. These results suggest that Dox induces inflammatory pyroptosis in Sol 8 cells, which is attenuated by ES-exos in vitro.

Survival and Differentiation of Adenovirus-Generated Induced Pluripotent Stem Cells Transplanted into the Rat Striatum

2014 ◽  
Vol 23 (11) ◽  
pp. 1407-1423 ◽  
Author(s):  
Kyle D. Fink ◽  
Julien Rossignol ◽  
Ming Lu ◽  
Xavier Lévêque ◽  
Travis D. Hulse ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Embryonic Stem ◽  
Tumor Formation ◽  
Rat Striatum ◽  
Tail Tip ◽  
Induced Pluripotent

Induced pluripotent stem cells (iPSCs) offer certain advantages over embryonic stem cells in cell replacement therapy for a variety of neurological disorders. However, reliable procedures, whereby transplanted iPSCs can survive and differentiate into functional neurons, without forming tumors, have yet to be devised. Currently, retroviral or lentiviral reprogramming methods are often used to reprogram somatic cells. Although the use of these viruses has proven to be effective, formation of tumors often results following in vivo transplantation, possibly due to the integration of the reprogramming genes. The goal of the current study was to develop a new approach, using an adenovirus for reprogramming cells, characterize the iPSCs in vitro, and test their safety, survivability, and ability to differentiate into region-appropriate neurons following transplantation into the rat brain. To this end, iPSCs were derived from bone marrow-derived mesenchymal stem cells and tail-tip fibroblasts using a single cassette lentivirus or a combination of adenoviruses. The reprogramming efficiency and levels of pluripotency were compared using immunocytochemistry, flow cytometry, and real-time polymerase chain reaction. Our data indicate that adenovirus-generated iPSCs from tail-tip fibroblasts are as efficient as the method we used for lentiviral reprogramming. All generated iPSCs were also capable of differentiating into neuronal-like cells in vitro. To test the in vivo survivability and the ability to differentiate into region-specific neurons in the absence of tumor formation, 400,000 of the iPSCs derived from tail-tip fibroblasts that were transfected with the adenovirus pair were transplanted into the striatum of adult, immune-competent rats. We observed that these iPSCs produced region-specific neuronal phenotypes, in the absence of tumor formation, at 90 days posttransplantation. These results suggest that adenovirus-generated iPSCs may provide a safe and viable means for neuronal replacement therapies.

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