K38 Keratinocyte Cell Line

The K38 Keratinocyte Cell Line has been established from the skin of neonatal WT mice. The prospective commercial application of the line is in the generation of in vitro epidermal ‘skin equivalents’ to be used as screening tools to look at the activity of NCE’s and/or NBE’s of dermatological interest, and/or to screen for such […]

Contributors

Inventor Institute
Julia Reichelt Newcastle University
Cat. #: 151682
Unit size: 1×10^6 cells / vial
Research Fields: Drug development
Organism: Mouse
Model: Primary line
Growth properties: Adherent
Product description: The K38 Keratinocyte Cell Line has been established from the skin of neonatal WT mice. The prospective commercial application of the line is in the generation of in vitro epidermal 'skin equivalents' to be used as screening tools to look at the activity of NCE's and/or NBE's 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 Murine keratinocyte stem cell line
Production details: Keratinocytes were isolated from neonatal wild-type BALB/c 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.
Cellosaurus ID: CVCL_W141
disease: Dermatology
Format: Frozen
Shipping conditions: Dry ice
Growth medium: The cell line is grown in Keratinocyte basal medium (low-calcium) (140116) supplemented with 10% FBS / FCS [Chelex 100 (Biorad)-treated to remove Ca2+ ions], 0.18 mM adenine, 0.5 μg/mL hydrocortisone, 5 μg/mL insulin,10 -10 M cholera toxin, 10 ng/mL EGF, 2 mM L-Glutamine, 1 mM pyruvate, 100 U/ml penicillin and 100 µg/ml streptomycin.
Subculture routine: Wash monolayer with PBS (without Ca2+ and Mg2+), incubate with EDTA solution (0.02% EDTA) for 5 min at room temperature and then incubate for 8-12 min with 0.05% trypsin/0.02% EDTA at 37°C until the cells come off easily. Add complete Keratinocyte medium and resuspend the cells by pipetting up and down 3-5 times. After centrifugation for 5 min at 220 x g, resuspend the pellet in complete Keratinocyte medium, and seed the cells at 3-5 x 10^4 cells/cm2 onto collagen I-coated plastic culture dish. Cultures are split 1:2 or 1:3, at confluency/2-3 times per week.
Temperature: 32°C
Atmosphere: 5% CO2
References:

Vollmers et al. 2012. Stem Cell Rev. 8(2):402-13. PMID: 21892602

Hoher et al. 2012. Stem Cell Rev. 8(2):426-34. PMID: 21874280

Reichelt et al. 2010. Methods Mol Biol. 585:59-69. PMID: 19907996

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