Mouse embryonic stem cell

Zhu, L. et al. (2007) DNA damage induced by multiwalled carbon nanotubes in mouse embryonic stem cells. Nano Letters Issues, 7 (12), 3592-3597. 

Fig. 13 Phase contrast microscope images of mouse embryonic stem cells in the PMBV/PVA hydrogel left) and on TCPS (right)...

Stavridis, M. P. and Smith, A. G. Neural differentiation of mouse embryonic stem cells. Biochem. Soc. Trans. 31 45-49, 2003. 

Kanellopoulou C, Muljo SA, Kung AL, Ganesan S, Drapkin R, Jenuwein T, Livingston DM, Rajewsky K (2005) Dicer-deficient mouse embryonic stem cells are defective in differentiation and centromeric silencing. Genes Dev 19 489—501... 

Gregory, R.I. et al. (2002) Inhibition of histone deacetylases alters allelic chromatin conformation at the imprinted U2afl-rsl locus in mouse embryonic stem cells. J. Biol. Chem. 277, 11728-11734. Thomas, G., Lange, H.W., and Hempel, K. (1975) Kinetics of histone methylation in vivo and its relation to the cell cycle in Ehrlich ascites tumor cells. Eur. J. Biochem. 51, 609-615. 

Sachinidis A, Eleischmann BK, Kolossov E, Wartenberg M, Sauer H, Hescheler J. Cardiac specific differentiation of mouse embryonic stem cells. Cardiovasc Res 2003 58 278-291. 

Bain G, Ray WJ, Yao M et al (1996) Retinoic acid promotes neural and represses mesodermal gene expression in mouse embryonic stem cells in culture. Biochem Biophys Res Commun 223(3) 691-694... 

NHEERL J1 mouse embryonic stem cells, 9-day (mESC) ACDC assay for cell number and... 

Barrier M, Jeffay S, Nichols HP, Chandler KJ, Hoopes MR, Slentz-Kesler K, Himter ES 111 (2011) Mouse embryonic stem cell adherent cell differentiation and cytotoxicity (ACDC) assay. Reprod Toxicol 31 383-391... 

The embryonic stem cell test is an animal-free alternative test method for developmental toxicity. Mouse embryonic stem cells are cultured in a hanging drop method to form embryoid bodies. These embryoid bodies, when plated on tissue culture dishes, differentiate to form contracting myocardial cell foci within 10 days. Inhibition of cardiomyocyte differentiation by test compounds serves as the end point of the assay, as monitored by cormting contracting muscle foci under the microscope. 

Pluripotent mouse embryonic stem cells (ES-D3 cell line, ATCC, Rockville, MD), Stock, with 1 x 10 cells/ml, stored in liquid Nitrogen. 

Fig. 1. Mouse embryonic stem cells (ES-D3) after passaging (a) and cultured for 2 days, ready for passaging (b). Unstained, light microscopy, magnification 4x.

Osman AM et al (2010) Proteome profiling of mouse embryonic stem cells to define markers for cell differentiation and embryo-toxicity. Reprod Toxicol 30 322-332. doi S0890-6238(10)00193-0(pii)10.1016/j. reprotox.2010.05.084... 

In 2007, the DART committee held a workshop on alternative assays, which was followed up by a workshop held at the European Teratology Society Annual Meeting in 2009. These workshops focused on three alternative assays (1) whole embryo culture (WEC), (2) mouse embryonic stem cell tests (mESC), and (3) zebrafish. Each assay was presented and data from users were shared, and strengths and limitations were discussed. It should be noted that the WEC and mESC are validated by ECVAM as alternative embryotoxicity assays. Still, there are numerous research needs before even validated tests can achieve regulatory acceptance. The discussions, conclusions, and recommendations of the 2007 workshop were published by Chapin et al. (14). Bullet lists of next steps to move forward were defined for each assay (14) and are briefly summarized here ... 

Functional analysis of mammalian genes by a large-scale gene trap approach in mouse embryonic stem cells... 

In their report the researchers describe a culturing technique that can turn mouse embryonic stem cells into cell clusters that resemble pancreatic islets. The clusters inner cells produced insulin, while outer cells produced glucagon and somatostatin, two other proteins typically synthesized by pancreatic cells. Most important, the embryonic stem cell-derived pancreas cells produce insulin in response to glucose, the fundamental role of beta cells that regulate insulin secretion. The major shortcoming of the system at this time is the low levels of insulin production. Refinements in culture technique or drug manipulation may be needed to achieve therapeutic levels. 

Vibrational imaging of living cells has been applied for studying intracellular lipid droplet organelles [86, 89, 90, 113, 114] (cf. Fig. 6.10A), monitoring receptor-mediated endocytosis [92], analyzing mouse embryonic stem cells... 

Doetschman, T., Gregg, R G., Maeda, N., Hooper, M. L., Melton, D. W., Thompson, S., et al. (1987) Targetted correction of a mutant HPRT gene in mouse embryonic stem cells. Nature 330, 576-578. 

Lei H, Oh SP, Okano M, Juttermann R, Goss KA, Jaenisch R, Li E. De novo DNA cytosine methyltransferase activities in mouse embryonic stem cells. Development 1996 122 3195-3205. 

Valancius, V. and Smithies, O. (1991) Testing of in-out targeting procedure for making subtle genomic modifications in mouse embryonic stem cells. Mol. Cell. Biol. 11, 1402-1408. 

Soto-Gutierrez A. 2007. Differentiation of mouse embryonic stem cells to hepatocyte-like cells by co-culture with human liver nonparenchymal cell lines. Nat. Protoc. 2, 347-356. 

The value of cultured stem cells to toxicology is in their promise as a continuous source of cells that can be induced to express a chosen cell-type-specific mature phenotype. This achievement would address two current limitations in safety testing scarcity of metabolically competent human systems and use of experimental animals. Numerous culture additives and conditions have been identified that cause commitment and partial differentiation along specific lineages, such as retinoic-acid-induced neuronal commitment of mouse embryonic stem cells. However, definition of conditions that produce fully differentiated cells with cell-type-specific function quantitatively similar to that of intact tissue remains problematic and is a high-priority research area in toxicology. 

In a recent study, scientists directed mouse embryonic stem cells to differentiate into DA neurons by introducing the gene Nurr 1. When transplanted into the brains of a rat model of Parkinson s disease, these stem cell-derived DA neurons reinnervated the brains of the mouse Parkinson s disease model, released dopamine, and improved motor function. 

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