GFP-KSC4 Keratinocyte stem cell line
11081701, mouse skin, Polygonal
Synonym(s):
KSC, KSC 4, KSC4
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product name
GFP-KSC4 Keratinocyte stem cell line, 11081701
biological source
mouse skin
growth mode
Adherent
karyotype
Not specified
morphology
Polygonal
receptors
Not specified
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Cell Line Origin
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 immortalized keratinocytes can be differentiated by the addition of calcium to the complete FAD medium at a final concentration of 1.2 mM (high calcium medium). Confluent cultures stratify when grown in high calcium medium.
DNA Profile
Not specified
Culture Medium
The cell line is grown in low calcium medium, DMEM/Ham′s F12 with FBS / FCS (F2442) 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 L-Glutamine (G7513), 1 mM pyruvate.
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 5 min at 220 g, 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.
Other Notes
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Disclaimer
This cell line has special release conditions: Commercial organisations are required to complete the ′Cell Line Release Authorisation for Research Use in Commercial Organisations′ release conditions form.
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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