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UK Health Security Agency

Mbd3KO

Mbd3KO

Catalogue No.

11041301

Cell Line Name

Mbd3KO

Cell Line Description

Mbd3KO stem cells have been genetically modified to produce a null phenotype for Mbd3, a critical determinant for lineage commitment in embryonic stem cells. This lack of Mbd3 (Mbd3 exons 2-7 have been replaced by an IRES-βgeo-pA sequence) allows them to be maintained in the absence of any exogenous factors and they fail to commit to developmental lineages. Upon re-introduction of the Mbd3 gene the cells are able to differentiate into identifiable bone, muscle, skin, fat and neuronal tissue. Mbd3 null embryonic stem cells can be used to study LIF-independent self-renewal to develop human stem cell lines and new cell culture media. They can also be used to develop specific stem cell lines for high throughput screening of therapeutic compounds. Finally they can be used to develop transgenics for the study of specific diseases or for the development of specific therapeutic stem cell lines for replacement therapies to treat certain diseases.

Characteristics

Receptors

Not specified

Products

Not specified

Tissue of Origin

Embryo

Morphology

Small spheroidal

Karyotype

Not specified

Disease

No associated disease stated

Culture Conditions

Cell Type

Stem cell

Subculture Routine

Plastic ware is pre-coated with gelatine as follows: porcine gelatine (Sigma G1890) is dissolved in sterile water (0.5g/500ml) at 50°C. The 0.1% solution is sterilized by filtration (0.22µm). Add 0.1% gelatine to plastic ware to cover bottom, and incubate for 20 minutes at room temperature. Remove gelatine, wash with PBS once and replace with appropriate culture medium. The flask/dish must not be allowed to dry out. An ampoule of Mbd3KO cells is thawed in 37°C water bath and the contents quickly transferred to a 15ml centrifuge tube. Medium is added drop wise to 5ml. Cells are centrifuged at 150 x g for 5 minutes. Cells are resuspended in 5ml of medium. Cells should be plated at 4-5 x 10⁴ cells/cm² using gelatine coated flasks. Cells must be subcultured every 2-3 days using 0.05% trypsin or trypsin/EDTA. Colonies must not be allowed to touch each other as overgrowth will result in differentiation. Cultures must be incubated in a humidified 5% CO₂/95% air incubator at 37°C.

Culture Medium

GMEM + 2mM Glutamine + 10% Foetal Bovine Serum (FBS) + 0.1 mM beta-mercaptoethanol + LIF 1000 Units/ml (ESGRO ESG1106). Use of LIF is optional, but its addition reduces differentiation.

Growth Mode

Adherent

Additional Info

Depositor

Edinburgh Research and Innovation Ltd, The University of Edinburgh, 1-7 Roxburgh street Edinburgh EH8 9TA

Country of Origin

United Kingdom

GMO Status

Genetically Modified Organism Class 1 (GMO1)

Hazard Group (ACDP)

Hazard Group (ACDP) 2

Applications

References

Kaji K, Caballero IM, MacLeod R, Nichols J, Wilson VA, Hendrich B. 2006 The NuRD component Mbd3 is required for pluripotency of embryonic stem cells. Nat Cell Biol. 8(3):285-92 PMID: 16462733.

Bibliography

Kaji K, Nichols J, Hendrich B. 2007 Mbd3, a component of the NuRD co-repressor complex, is required for development of pluripotent cells. Development. 134(6):1123-32 PMID: 17287250.

Available Formats

  • Frozen
  • DNA-5µg (100ng/µl)

If use of this culture results in a scientific publication, it should be cited in the publication as: Mbd3KO (ECACC 11041301).

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.

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.