Effects of DMSO on the Pluripotency of Cultured Mouse Embryonic Stem Cells (mESCs)

DMSO is a commonly used solvent in biological studies, as it is an amphipathic molecule soluble in both aqueous and organic media. For that reason, it is the vehicle of choice for several water-insoluble substances used in research. At the molecular and cellular level, DMSO is a hydrogen-bound disrupter, an intercellular electrical uncoupler, and a cryoprotectant, among other properties. Importantly, DMSO often has overlooked side effects. In stem cell research, the literature is scarce, but there are reports on the effect of DMSO in human embryoid body differentiation and on human pluripotent stem cell priming towards differentiation, via modulation of cell cycle. However, in mouse embryonic stem cell (mESC) culture, there is almost no available information. Taking into consideration the almost ubiquitous use of DMSO in experiments involving mESCs, we aimed to understand the effect of very low doses of DMSO (0.0001%-0.2%), usually used to introduce pharmacological inhibitors/modulators, in mESCs cultured in two different media (2i and FBS-based media). Our results show that in the E14Tg2a mESC line used in this study, even the smallest concentration of DMSO had minor effects on the total number of cells in serum-cultured mESCs. However, these effects could not be explained by alterations in cell cycle or apoptosis. Furthermore, DMSO did not affect pluripotency or differentiation potential. All things considered, and although control experiments should be carried out in each cell line that is used, it is reasonable to conclude that DMSO at the concentrations used here has a minimal effect on this particular mESC line.

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Mouse embryonic stem cell differentiation into cardiomyocytes is inhibited by ethanol but only after the embryoid body stage (844.3)

The FASEB Journal ◽

10.1096/fasebj.28.1_supplement.844.3 ◽

2014 ◽

Vol 28 (S1) ◽

Author(s):

Gregory DeKrey ◽

Sarah Worley Jones ◽

Alex Terry ◽

Catherine Gardiner ◽

Brittney Vaughn

Keyword(s):

Stem Cell ◽

Cell Differentiation ◽

Embryonic Stem Cell ◽

Embryoid Body ◽

Embryonic Stem ◽

Mouse Embryonic Stem Cell ◽

Stem Cell Differentiation ◽

Embryonic Stem Cell Differentiation

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Embryoid-body cells derived from a mouse embryonic stem cell line show differentiation into functional hepatocytes

Hepatology ◽

10.1053/jhep.2002.34136 ◽

2002 ◽

Vol 36 (1) ◽

pp. 22-29 ◽

Cited By ~ 192

Author(s):

Ryoko Chinzei ◽

Yujiro Tanaka ◽

Keiko Shimizu-Saito ◽

Yuzuru Hara ◽

Sei Kakinuma ◽

...

Keyword(s):

Stem Cell ◽

Embryonic Stem Cell ◽

Cell Line ◽

Embryoid Body ◽

Embryonic Stem Cell Line ◽

Embryonic Stem ◽

Mouse Embryonic Stem Cell ◽

Stem Cell Line

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HoxA2 Regulates Proliferation of an Embryonic Megakaryocyte Progenitor, Which Can Effectively Produce Platelets In Vitro.

Blood ◽

10.1182/blood.v110.11.1266.1266 ◽

2007 ◽

Vol 110 (11) ◽

pp. 1266-1266

Author(s):

Michelina Iacovino ◽

Mitsujiro Osawa ◽

Zhaohui Xu ◽

Melissa G. Prather ◽

Michael Kyba

Keyword(s):

Bone Marrow ◽

Hox Genes ◽

Embryoid Body ◽

Embryonic Stem Cell Line ◽

Embryonic Stem ◽

Mouse Embryonic Stem Cell ◽

Rare Cells ◽

Embryoid Body Differentiation ◽

Megakaryocyte Progenitors

Abstract Megakaryocytes are rare cells in the bone marrow, and are solely responsible for thrombopoiesis throughout the life of the organism. Various transcription factors regulating megakaryopoiesis have been identified, including GATA1, Fli1 and NF-E2, which are important for the maturation and differentiation of megakaryocytes and SCL and FOG1, which play important roles in the specification and proliferation of the early megakaryocyte progenitor but no single factor capable of sustaining long-term proliferation of the megakaryocyte progenitor has been identified. The rarity of this progenitor, and the difficulty of proliferating megakaryocytes in vitro have seriously hampered the effort to produce large quantities platelets in vitro, and to date the only source of platelets for transplantation remains human donors. In this study, through a gain of function screen of Hox genes in which each paralog group was tested, we identified HoxA2 as an important transcription factor for the proliferation of early embryonic megakaryocytic progenitors. HoxA2 promoted the outgrowth of undifferentiated megakaryocyte progenitors when transduced into precirculation yolk sac stem and progenitor cells, however it had no effect on megakaryocytic progenitors derived from definitive HSCs of adult bone marrow. To study this unique embryonic regulatory circuit further, we generated an inducible mouse embryonic stem cell line, in which the expression of murine HoxA2 is regulated by doxycycline. Embryoid body differentiation of these cells, followed by sorting and reculture of the Kit+/CD41+ hematopoietic progenitor fraction at day 6 in the presence of continual HoxA2 expression allowed undifferentiated c-Kit+/CD41+/CD45+ megakaryocyte progenitors to proliferate for upwards of 2 months. When doxycycline is removed, numerous gene expression changes take plase, and these progenitors differentiate into mature c-Kitneg/CD41high/CD45neg megakaryocytes, which undergo proplatelet formation, releasing large quantities of platelets (∼107/ml) into the medium. As this rate of production can be sustained for weeks, a highly efficient platelet bioreactor is enabled. The platelets express the integrin αIIb-β3 (GPIIb-IIIa complex), and upon stimulation they bind fibrinogen, demonstrating functionality. We are currently investigating the transplantation potential of these cells.Our data suggest that adult and embryonic megakaryopoiesis have distinct regulatory circuits, and demonstrate a novel and uniquely embryonic role for HoxA2, which opens up the possibility of producing industrial quantities of platelets in vitro for therapeutic purposes.

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Shear-Controlled Single-Step Mouse Embryonic Stem Cell Expansion and Embryoid Body-Based Differentiation

Stem Cells ◽

10.1634/stemcells.2005-0112 ◽

2005 ◽

Vol 23 (9) ◽

pp. 1333-1342 ◽

Cited By ~ 187

Author(s):

Elaine Y.L. Fok ◽

Peter W. Zandstra

Keyword(s):

Stem Cell ◽

Embryonic Stem Cell ◽

Cell Expansion ◽

Embryoid Body ◽

Embryonic Stem ◽

Mouse Embryonic Stem Cell ◽

Single Step ◽

Stem Cell Expansion

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Characterization of Liposarcoma Cell Lines for Preclinical and Biological Studies

Sarcoma ◽

10.1155/2012/148614 ◽

2012 ◽

Vol 2012 ◽

pp. 1-9 ◽

Cited By ~ 17

Author(s):

Eva W. Stratford ◽

Russell Castro ◽

Jeanette Daffinrud ◽

Magne Skårn ◽

Silje Lauvrak ◽

...

Keyword(s):

Stem Cell ◽

Cell Lines ◽

Metastatic Potential ◽

Cellular Level ◽

Differentiation Markers ◽

Differentiation Potential ◽

Biological Studies ◽

Novel Drugs

Liposarcoma cell lines representin vitromodels for studying disease mechanisms at the cellular level and for preclinical evaluation of novel drugs. To date there are a limited number of well-characterized models available. In this study, nine immortal liposarcoma cell lines were evaluated for tumor-forming ability, stem cell- and differentiation potential, and metastatic potential, with the aim to generate a well-characterized liposarcoma cell line panel. Detailed stem cell and differentiation marker analyses were also performed. Five of the liposarcoma cell lines were tumorigenic, forming tumors in mice. Interestingly, tumor-forming ability correlated with high proliferative capacityin vitro. All the cell lines underwent adipocytic differentiation, but the degree varied. Surprisingly, the expression of stem cell and differentiation markers did not correlate well with function. Overall, the panel contains cell lines suited forin vivoanalyses (LPS141, SA-4, T778, SW872, and LISA-2), for testing novel drugs targeting cancer stem cells (LPS141) and for studying tumor progression and metastasis (T449 and T778).

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