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UK Health Security Agency

COCA

COCA

Catalogue No.

10112001

Cell Line Name

COCA

Cell Line Description

COCA, a murine epidermal cell line was developed by serially passaging keratinocytes from the back skin of adult C57BL/DBA mice. Culture of the COCA cells is in fully defined media without requirement for feeder layer or other coating. COCA retains its ability to differentiate and stratify in response to increased calcium concentrations thus providing an excellent experimental system for in vitro (conventional and 3D epidermal cell cultures) and in vivo (skin regeneration) skin modelling.

Discontinued Message

Cell line temporarily discontinued pending investigation.

Characteristics

Receptors

Not specified

Products

See Segrelles et al 2011

Tissue of Origin

Epidermis

Karyotype

See Segrelles et al 2011

Disease

None Stated

Culture Conditions

Cell Type

Epithelial

Subculture Routine

A centrifugation step to remove the cryoprotectant is essential. Rapidly thaw the frozen ampoule in a water bath at 37°C for 1-2 minutes. Transfer the contents to a centrifuge tube and slowly add 5-10ml of pre-warmed growth media. Remove a sample for counting. Centrifuge at 100g for 2-3 minutes to pellet cells and seed at approximately 1.5 x 104 cells/cm².

Split sub-confluent cultures (70-80%) 1:3 to 1:6 i.e. seeding at 0.5-1.5 x 104 cells/cm² using 0.05% trypsin or trypsin/EDTA. It is important to inactivate the trypsin with an equal volume of trypsin inhibitor. To remove all traces of trypsin and trypsin inhibitor pellet the cells by centrifugation and resuspend the cells in fresh medium. Media change every second day.

Recommended Growth conditions: 5% CO2; 37°C

Culture Medium

CnT-07 media (CELLnTEC, Bern, Switzerland. http://www.cellntec.com/products/epidermal/culture-media/cnt-07) This defined medium contains low calcium (0.07mM). To promote good cell attachment post trypsinisation, an 8-14 hours incubation in CnT-07 medium containing 0.2mM CaCl₂ is necessary. Differentiation can be initiated by transferring a confluent monolayer from 0.07mM calcium to high calcium (1.2mM). In Segrelles et al 2011 PMID: 21510892, 3D in vitro cultures were performed using CnT-02-3D cell culture media from CELLnTEC (Bern, Switzerland). The company has recently changed the formulation of this culture media to CnT-02-3D Prime (CnT-02-3DP) media. The ability of both media (CnT-02-3D Classic the original vs. Prime) to grow 3D cultures using COCA keratinocytes has been determined by the authors. They found that (see attachment below) the recently developed Cnt-02-3D Prime media is not efficient to sustain 3D culture of COCA cells. CELLnTEC have confirmed that they will produce and provide CnT-02-3D Classic when requested.

Growth Mode

Adherent

Additional Info

Depositor

Dr. Corina Lorz, Molecular Oncology Unit, Division of Biomedicine, Department of Basic Research, CIEMAT (ed 70A), Ave Complutense 22; 28040 Madrid, Spain.

Country of Origin

Spain

Hazard Group (ACDP)

Hazard Group (ACDP) 2

Applications

References

Carmen Segrelles, Almudena Holguin, Pilar Hernandez, José M Ariza, Jesus M Paramio and Corina Lorz. 2011 Establishment of a murine epidermal cell line suitable for in vitro and in vivo skin modelling. BMC Dermatology Apr 21; 11(1):9 PMID: 21510892.

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: COCA (ECACC 10112001).

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.