Catalogue No.
16010601
Nd ES
Nd ES
Cell Line Name
Nd ES
Cell Line Description
A mouse embryonic stem cell (ES) line (Nd) containing a novel fluorescent reporter (VNP) to investigate the temporal dynamics of Nanog expression in mouse and to dissect the lineage potential at different Nanog expression levels. This novel reporter ES cell line allows the accurate monitoring the dynamic expression of the transcription factor Nanog in the pluripotent state. The cell line will be a valuable tool to obtain quantitative measurements of global gene expression in pluripotent mouse ES cells in different states, allowing a detailed molecular mapping of the pluripotent state.
General Info
Species
Mouse
Links
Cellosaurus: CVCL_IU77
Release Conditions
Characteristics
Tissue of Origin
Embryo
Morphology
Small round cell growing in spherical colonies
Applications
Monitoring Nanog protein heterogeneity and expression dynamics in mouse embryonic stem cells in pluripotent state and during early differentiation. Formation of embryoid bodies consisting of three germ layers ectoderm, mesoderm and endoderm as assessed by expression of lineage markers Fgf5, T-brachyury and Gata6, respectively. Formation of typical neuroepithelial rosette-like structures when cultured in monolayer in the absence of Leukemia Inhibitory Factor (LIF)
Disease
None Stated
Culture Conditions
Cell Type
Stem cell
Subculture Routine
Cultures must be incubated in a humidified 10% CO₂/95% air incubator at 37°C. Subculture with 0.05% Trypsin/EDTA and inoculate new cultures at 3 x 10⁴ per cm².
Culture Medium
Culture cells in Glasgow Modified Eagles Medium with 10% FCS + 2mM L-glutamine, 2ng/ml Leukemia Inhibitory Factor (LIF) and 1mM 2-mercaptoethanol on gelatin-coated (0.1%) cell culture plastic.Alternatively, ES cells can be cultured in 2i medium (iStem medium, Stem Cells, Inc) supplemented with 2ng/ml LIF and inhibitors of the ERK pathway and of GSK3β (Ying QL et.al 2008)
Growth Mode
Adherent
Additional Info
Depositor
Ana MV Guedes
Country of Origin
Portugal
Genetic Modification
Yes. Nd cells are a novel ES cell line containing a transgenic bacterial artificial chromosome (BAC) with a fluorescent reporter (yellow fluorescent protein, Venus) inserted under the control of the Nanog regulatory regions.
Hazard Group (ACDP)
Hazard Group (ACDP) 2
Applications
References
Abranches E, Bekman E, Henrique D. Generation and characterization of a novel mouse embryonic stem cell line with a dynamic reporter of Nanog expression. PLoS One. 2013; 8(3):e59928. doi: 10.1371/journal.pone.0059928. Epub 2013 Mar 19. Erratum in: PLoS One. 2014;9 (6):e101028. PubMed PMID: 23527287; PubMed Central
Bibliography
Ying QL, Wray J, Nichols J, Batlle-Morera L, Doble B, Woodgett J, Cohen P, Smith A (2008) The ground state of embryonic stem cell self-renewal. Nature 453:519-523 Nair G, Abranches E, Guedes AM, Henrique D, Raj A. Heterogeneous lineage marker expression in naive embryonic stem cells is mostly due to spontaneous differentiation. Sci Rep. 2015 Aug 21; 5: 13339. doi: 10.1038/srep13339. PMCID: PMC4544010. PMID: 26292941 Sanchez-Ripoll Y, Bone HK, Owen T, Guedes AM, Abranches E, Kumpfmueller B, Spriggs RV, Henrique D, Welham MJ.Glycogen synthase kinase-3 inhibition enhances translation of pluripotency-associated transcription factors to contribute to maintenance of mouse embryonic stem cell self-renewal. PLoS One. 2013; 8 (4):e60148. doi: 10.1371/journal.pone.0060148. Epub 2013 Apr 5. PMCID: PMC3618116. PMID: 23577087.
Available Formats
- Frozen
- DNA-5µg (100ng/µl)
Citation Guidance
If use of this culture results in a scientific publication, it should be cited in the publication as: Nd ES (ECACC 16010601).
Biosafety Information
Unless specified otherwise, at ECACC we routinely handle all of our cell lines at containment level 2 in accordance with the ACDP guidelines (Advisory Committee on Dangerous Pathogens) (UK). All cell cultures have the potential to carry as yet unidentified adventitious agents. It is the responsibility of the end user to ensure that their facilities comply with biosafety regulations for their own country.
ACDP Guidance: Biological agents: Managing the risks in laboratories and healthcare premises.
Further Information
The Culture Collections represent deposits of cultures from world-wide sources. While every effort is made to ensure details distributed by Culture Collections are accurate, Culture Collections cannot be held responsible for any inaccuracies in the data supplied. References where quoted are mainly attributed to the establishment of the cell culture and not for any specific property of the cell line, therefore further references should be obtained regarding cell culture characteristics. Passage numbers where given act only as a guide and Culture Collections does not guarantee the passage number stated will be the passage number received by the customer.
Cultures supplied by Culture Collections are for research purposes only. Enquiries regarding the commercial use of a cell line are referred to the depositor of the cell line. Some cell lines have additional special release conditions such as the requirement for a material transfer agreement to be completed by the potential recipient prior to the supply of the cell line. Please view the Terms & Conditions of Supply for more information.