Improved barrier as seen by raman spectroscopy penetration profiles.
The 3D Starter Kit with Primary Keratinocytes (PR3D-HPEK-50) contains everything needed to establish 3D epidermal models with improved differentiation and barrier function in your own lab.
The kit contains primary cells, CnT-PR proliferation mediun, CnT-PR-3D Barrier Medium, and (effective July 2014) 48 inserts for air-lift culture. For details, please see the datasheet.
The 3D models can be routinely established using our recommended protocol
Following this protocol, the 3D models deliver up to a 50% better barrier function than traditional media. More detailed raman spectroscopy results can be found in the following link:
Improved Barrier Function with 3D Barrier Medium
3D Keratinocyte Starter Kit including Primary Keratinocytes
3D keratinocyte starter Kit with Primary Keratinocytes provides all components required for the establishment of 3 dimensional keratinocyte cultures with improved barrier function. The kit includes primary human keratinocytes, CnT-Prime Medium (CnT-PR) for monolayer culture, CnT-Prime 3D Barrier Medium (CnT-PR-3D), and 48 cell culture inserts (0.4 um pore size). Optional extra: CnT-ST-100 stain solution for evaluating confluency prior to airlift.
The cells are of human origin and should therefore be handled as hazardous. Although it has been tested negative for HIV 1, Hepatitis B and Hepatitis C, treat this material as potentially infectious, and use appropriate biocontainment, protective equipment and other precautions to prevent accidental exposure.
- HPEKp Keratinocytes: Human, pooled (3 or more donors), juvenile.
- Tissue type
- Pack size
- PR3D-HPEK-50 is a kit consisting of:
- 1 x HPEKp: Human Epidermal Keratinocyte Progenitors > 5 x 105 viable cells per vial (1 mL)
- 1 x CnT-PR: 500 mL kit of CnT-Prime Epithelial Culture Medium
- 1 x CnT-PR-3D: one 500 mL kit of CnT-Prime 3D Barrier Medium
- 48 x Cell culture inserts: polycarbonate membrane, 12 mm diameter, 0.4 µm pore size
- 1 x PR3D-HPEK-50 Data sheet
- 1 x PR3D-HPEK-50 Protocol
- Culture medium
- Media included in the kit: CnT-PR Epithelial Medium, CnT-PR-3D 3D Barrier Medium
All media are designed for use in a 5% CO2 atmosphere.
Media must be thawed just prior to use. If lipids in 3D Barrier medium do not resolubilise in cold medium after thawing, they will be visible as a faint white layer in the bottom of the bottle. To solubilise, warm bottle to room temperature, and solubilise with gentle swirling.
- For thawing, seeding and passaging instructions, please see the general cultivation protocol in the resources section of www.cellntec.com
- Recommended seeding density after passaging: 4 x 103 cells / cm2. For passaging instructions, please see also general cultivation protocol, on our resources section.
- Av. time to confluence
- 5 to 7 days for monolayer culture of keratinocytes (depending on temperature, seeding density and protocol)
- Keratinocytes guaranteed to provide 20 population doublings when grown in CnT-Prime medium. For optimal 3D stratification, use cells within 12 population doublings of thawing.
- Storage / Shelf life
- Immediately upon arrival transfer the cryo vial to the liquid nitrogen container, until ready to use. Medium storage: -20°C. Thaw overnight at 4oC prior to use, or alternatively at room temperature for approx 5 hours. Do not thaw in a hot water bath. For expiry date of frozen medium, refer to medium label. Storage of thawed medium: 4°C in the dark, use within 6 weeks of thawing. Important: The quality of cell culture media is rapidly degraded by light. Store in the dark, and minimize all light exposure during handling. Inserts: room temperature. One year from date of purchase.
- Recommended seeding density after thawing: 2 to 4 x 103 cells / cm2. For thawing instructions, please see also general cultivation protocol, on the resources section of www.cellntec.com
- Recommended freezing density: 1 x 106 cells / mL. For freezing instructions for keratinocytes, please see also general cultivation protocol, on the resources section of www.cellntec.com
- Quality control
- Primary cells tested negative for Hepatitis B, Hepatitis C and HIV-1.
Media tested free of bacteria, fungi and mycoplasma contamination.
- Shipping condition
- All kit components are shipped together on dry ice.
- All kit components can be purchased separately.
Optional extra available separately: CnT-ST-100 for evaluating confluence prior to air-lift.
- Intended use
- For research use only. Not for use in therapy or diagnostics.
- Last update