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UK Health Security Agency

LF2

LF2

Catalogue No.

16022532

Cell Line Name

LF2

Cell Line Description

The LF2 mouse embryonic stem cell line is a germ-line competent embryonic stem cell line that inactivates one of its X chromosomes upon retinoic acid treatment.

General Info

Species

Mouse: female Mus musculus, strain 129/Ola

Unique to ECACC

Yes

Release Conditions

Restricted - commercial organisations are required to complete the 'Cell Line Release Authorisation for Research Use in Commercial Organisations' release conditions form in the supporting documents section.

Characteristics

Tissue of Origin

Embryo

Morphology

Small round cell growing in larger spherical colonies

Applications

Differentiation of LF2 cells can be induced by removing LIF and by using 100 nM all-trans-retinoic acid in DMEM supplemented with 10% foetal bovine serum and 10-4 mM 2-mercaptoethanol The differentiation medium should be changed daily. All cells grown at 37°C in 8% CO₂. Embryonic bodies are formed by growing cells in low adherence dishes in DMEM with FBS for 10 days. Embryonic stem cells provide a powerful system for the experimental analysis of mammalian development. Their genetic manipulation allows the introduction of defined modifications into the mouse germ-line, whilst in culture they constitute both an assay system for, and a potential source of, embryonic regulatory factors.

Disease

None Stated

Culture Conditions

Cell Type

Stem cell

Subculture Routine

Split sub-confluent cultures (70-80%) 1:3 to 1:6 i.e. seeding at 2-4x10⁴ cells/cm² using 0.05% trypsin/EDTA solution. Cells are cultured at 37°C, in 95% air and 5% CO₂, and passaged every 3 to 4 days with a media change daily. Culture cells on gelatin-coated flasks or plates (left to dry for a minimum of 2 hours before use) and pre-seed with Mitromycin-C treated mouse embryonic fibroblasts Feeder layers are prepared in the flasks at least 24 hours in advance of being required. When cultures of treated mouse embryonic fibroblasts have been prepared, an ampoule is thawed in 37°C water bath and the contents quickly transferred to a 15ml centrifuge tube containing LF2 growth medium and cells seeded at 1-3 x 10⁴ cells/cm². These cells grow rapidly and it is recommended media be changed daily.

Culture Medium

ES cell line LF2 was grown on gelatin-coated flasks or plates and pre-seeded with lethally irrediated or Mitomycin-C-treated mouse embryonic fibroblasts. Maintain LF2 cells in an undifferentiated state in DMEM with GlutMAX, 15% foetal calf serum (Gibco), 10-4 mM 2-mercaptoethanol and 1,000 U/ml of Leukemia Inhibitory Factor (LIF).

Growth Mode

Adherent

Additional Info

Depositor

Ximbio (formerly known as Cancer Research Technologies Limited)

Country of Origin

United Kingdom

Hazard Group (ACDP)

Hazard Group (ACDP) 2

Applications

References

Peng et al. 2009. Cell. 139(7):1290-302. PMID: 20064375. Marks et al. 2009. Genome Res. 19(8):1361-73. PMID: 19581487. Kabuyama et al. 2007. Genes Cells. 12(11):1235-44. PMID: 17986007. Bernstein et al. 2006. Mol Cell Biol. 26(7):2560-9. PMID: 16537902. Hake et al. 2006. J Biol Chem. 281(1):559-68. PMID: 16267050. Atamas et al. 2003. Am J Respir Cell Mol Biol. 29(6):743-9. PMID: 12805086. Heard et al. 2001. Cell. 107(6):727-38. PMID: 11747809. Mermoud et al. 1999. J Cell Biol. 147(7):1399-408. PMID: 10613899. Keohane et al. 1996. Dev Biol. 180(2):618-30. PMID: 8954732. Chan et al. 1991. J Exp Med. 173(4):869-79. PMID: 1672545. Nichols et al. 1990. Development. 110(4):1341-8. PMID: 2129226 Rougeulle C et al. 2004 Mol Cell Biol 24: 5475-5484 PMID: 15169908

Bibliography

Smith, A. G. 1991. Culture and differentiation of embryonic stem cells. J. Tissue. Cult. Methods 13:89-94.

Available Formats

  • Frozen

If use of this culture results in a scientific publication, it should be cited in the publication as: LF2 (ECACC 16022532).

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.