Improved barrier as seen by raman spectroscopy penetration profiles.
The 3D Starter Kit without Primary Keratinocytes (PR3D-K-50) contains the media and inserts needed to establish 3D epidermal models with improved differentiation and barrier function in your own lab.
Early passage human keratinocytes growing in CnT-PR medium are also required
The kit contains 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, without Primary Keratinocytes
3D keratinocyte starter Kit was developed for the establishment of 3 dimensional keratinocyte cultures with improved barrier function. Kit includes 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). Kit does not contain primary keratinocytes. Optional extra: CnT-ST-100 stain solution to evaluate 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.
- Tissue type
- Pack size
- PR3D-K-50 is a kit consisting of:
- 1 x CnT-PR: 500 mL kit of CnT-Prime Epithelial Culture Medium
- 1 x CnT-PR-3D: one 500 mL kit of CnT-PR-3D 3D Barrier Medium
- 48 x Cell culture inserts: polycarbonate membrane, 12 mm diameter, 0.4 µm pore size
- 1 x PR3D-K-50 Data sheet
- 1 x PR3D-K-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.
- To establish the 3D epidermal models, early passage primary keratinocytes growing strongly in CnT-Prime medium are required. For recommended keratinocyte isolation, 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 growing in CnT-Prime medium (depending on temperature, seeding density and protocol)
- Storage / Shelf life
- Medium storage: -15°C or below. Thaw overnight at 4oC prior to use, or alternatively at room temperature for approx. 5 hours. Do not thaw media in 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. CnT-ST: room temperature. One year from date of purchase.
- Quality control
- Media tested free of bacteria, fungi and mycoplasma contamination.
- Shipping condition
- All kit components are together on dry ice.
- HPEKp primary keratinocytes and all culture media can also be purchased individually. CnT-ST-100 stain solution for evaluation of confluency prior to airlift is also available separately.
- Intended use
- For research use only. Not for use in therapy or diagnostics.
- Last update