|Supplied by:||European Collection of Authenticated Cell Cultures (ECACC)|
|Culture Type:||Cell line|
|Collection:||ECACC General Collection|
|Cell Line Name:||GFP-KSC4 Keratinocyte stem cell line|
|Citation Guidance:||If use of this culture results in a scientific publication, it should be cited in the publication as: GFP-KSC4 Keratinocyte stem cell line (ECACC 11081701)|
|Keywords:||Murine keratinocyte stem cell line, genetically modified,|
|Cell Line Description:||Keratinocytes were isolated from neonatal homozygous C57BL/6 Tg(CAG EGFP)1Osb/J mouse skin. The cell line was established by growing cells for the first 8 passages in co culture with 3T3 J2 fibroblast feeder cells. From passage 9 onwards cells were grown without feeder cells. This line can be used for the generation of in vitro epidermal skin equivalents to be used as screening tools to look at the activity of NCEs and/or NBEs of dermatological interest, and/or to screen for such NCE and/or NBE in high throughput systems. It may also be of use to non pharmaceutical healthcare and/or cosmetic companies involved in the personal vitality and/or beauty product development segments.The spontaneously immortalised keratinocytes can be differentiated by the addition of calcium to the complete FAD medium at a final concentration of 1.2mM (high calcium medium). Confluent cultures stratify when grown in high calcium medium.|
|Tissue of Origin:||skin|
|GMO Status:||Genetically Modified Organism Class 1 (GMO1)|
Unless specified otherwise, at the European Collection of Authenticated Cell Cultures (ECACC) we routinely handle all of our cell lines at containment level 2 in accordance with the ACDP guidelines. ACDP = 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.
Hyperlinks to MSDS documents:
Frozen cell cultures Material Safety Data Sheet
Growing cell cultures Material Safety Data Sheet
Nucleic acids derived from cell cultures Material Safety Data Sheet
|Subculture Routine:||Wash monolayer with PBS (without Ca2+ and Mg2+), incubate with EDTA solution for 5 min at room temperature and finally incubate for 8-12 min with 0.05% trypsin/0.02% EDTA at 37°C until they come off easily. Add complete FAD medium and resuspend the cells by pipetting up and down 3-5 times. After centrifugation for 5min at 220g resuspend the pellet in complete FAD medium and seed the cells onto collagen I-coated dishes. Seed new cultures at 1.5-3 x 104 cells/cm2.|
|Culture Medium:||The cell line is grown in low calcium medium, DMEM/Ham's F12 with 10% FCS and several supplements at 32°C, 5% CO2. The supplements are: 0.18 mM adenine, 0.5 µg/ml hydrocortisone, 5 µg/ml insulin,10 -10 M cholera toxin, 10 ng/ml EGF, 2 mM, glutamine, 1 mM pyruvate. For detailed instructions please see attachment below.|
|Depositor:||Cancer Research Technology Ltd Angel Building 407 St John Street London EC1V 4AD. Originator: Dr J Reichelt, Institute of Cellular Medicine, Framlington Place, Newcastle University, Newcastle upon Tyne, NE2 4HH|
|References:||Reichelt J, Haase I. 2010 Establishment of spontaneously immortalized keratinocyte lines from wild-type and mutant mice. Methods Mol Biol.585:59-69.PMID: 19907996|
|Additional Bibliography:||Okabe M, Ikawa M, Kominami K, Nakanishi T, Nishimune Y 1997 'Green mice' as a source of ubiquitous green cells. FEBS Lett.407(3):313-9.PMID: 9175875 Vollmers A, Wallace L, Fullard N, Höher T, Alexander MD, Reichelt J 2011 Two- and Three-Dimensional Culture of Keratinocyte Stem and Precursor Cells Derived from Primary Murine Epidermal Cultures. Stem Cell Rev.Sep 3. [Epub ahead of print] PMID: 21892602|
|Patents:||None specified by Depositor|
|Release Conditions:||Yes - commercial organisations are required to complete the 'Cell Line Release Authorisation for Research Use in Commercial Organisations' release conditions form.|
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.
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