- New Orders: MatTek Scientists will consult on your application at no additional charge. It is STRONGLY RECOMMENDED that you have this consultation BEFORE placing your first order to help ensure that all media and accessories needed to perform your application are included in the initial order.
- ALL Tissue Orders: Please allow 3-4 weeks from order date to delivery of tissues. Please contact MatTek Customer Service for additional details.
- Type: Normal human oral keratinocytes (NHOK) are differentiated into tissues with a non-cornified, buccal phenotype, part number ORL-200 (replaced ORL-100) or a cornified, gingival phenotype, part number GIN-100. The same cells are used in both tissues (differentiation conditions differ).
- Genetic make-up: Single donor.
- Derived from: Non-diseased, adult human oral tissue obtained from patients undergoing tooth extractions.
- Screened for: HIV, Hepatitis-B, Hepatitis-C, mycoplasma, bacteria, yeast, fungi.
- Base medium: Dulbecco's Modified Eagle's Medium (DMEM).
- Growth factors/hormones: Epidermal growth factor and other proprietary factors.
- Serum: None.
- Antibiotics: Gentamicin 5 µg/ml (10% of normal gentamicin level).
- Anti-fungal agent: Amphotericin B 0.25 µg/ml.
- pH Indicator: Phenol red.
- Other additives: Proprietary.
- Alternatives: Phenol red-free, antibiotic-free, anti-fungal-free medium and tissue are available. Agents are removed 3 days prior to shipment.
- Maintenance medium: ORL-200-MM or GIN-100-MM.
- Kits: The EpiOral kits (ORL-200 or ORL-100-IB) and EpiGingival kits (GIN-100) consist of 24 tissues. ORL-606 and GIN-606 kits consist of 6 tissues. (Tissue "kits" contain tissues, a small amount of culture medium, and plasticware; contact MatTek for specific kit contents.)
- Substrate: ORL-200: Collagen coated, 9 mm ID single well tissue culture plate inserts are used - Millipore Millicell™ CM single well tissue culture plate inserts. Pore size = 0.4 µm, Inner Diameter = 0.875 cm. Surface area = 0.6 cm2.
ORL-606: Millipore Millicell™ CM single well tissue culture plate inserts. Pore size = 0.4 µm, Inner Diameter = 2.5 cm. Surface area = 4.2 cm2.
- Culture: At air liquid interface.
- Histology: ORL-200: 8-11 cell layers of non-cornified tissue. GIN-100: 8-11 cell layers of cornified tissue.
- Lot Numbers: Tissue lots produced by each technician for each week are assigned a specific lot number. Typically, there are multiple lot numbers for any given week's tissue production. A letter of the alphabet is appended to the end of the lot number to differentiate between individual kits within a given lot of tissues. All tissue kits within a lot are identical with regard to cells, medium, handling, culture conditions, etc.
- Shipment: At 4°C on medium-supplemented, agarose gels.
- Shipment day: Monday.
- Delivery: Tuesday morning via FedEx priority service (US). Outside US: Tuesday-Thursday depending on location.
- Shelf life: Including time in transit, tissues may be stored at 4°C for up to 4 days prior to use. However, extended storage periods are not recommended unless absolutely necessary. In addition, the best reproducibility will be obtained if tissues are used consistently on the same day, e.g. Wednesday morning following overnight storage at 4°C.
- Length of experiments: Cultures can be continued for at least 1 week with good retention of normal morphology. Cultures must be fed every other day with 5.0 ml of maintenance medium (ORL-200-MM or GIN-100-MM). Cell culture inserts are placed atop culture stands (MEL-STND; click on this link for photo) or washers (EPI-WSHR) in 6-well plates to allow the use of 5.0 ml.
- Alternative tissues:
GIN-100: Similar to ORL-200 except tissues are cornified at the apical surface.
ORL-100-IB: Similar to ORL-200 except tissue is 20-30 cell layers thick.
ORL-606: Similar to ORL-200 except 6 tissues per kit, each tissue cultured in a larger diameter tissue culture insert.
ORL-300-FT: Full thickness ORL-200.
ORL-300-FT-LC: Full thickness ORL-200 with Langerhans Cells.
V. Quality Control & Sterility
- Visual inspection: All tissues are visually inspected and if physical imperfections are noted, tissues are rejected for shipment.
- End-use testing: Transepithelial electrical resistance (TEER) of each lot is measured prior to shipment (QC specification: ORL-200: 413 + 138 ohms*cm2; GIN-100: 516 + 122 ohms*cm2). Tissues are also exposed to 1.0% Triton X-100 for 60, 120 and 240 minutes. The time of exposure required to reduce the tissue viability (ET-50) using the MTT assay is determined (QC specification: ORL-200: 61.0 + 19.8 min.; GIN-100: 7.97 + 0.8 hrs.). A summary of these results will be provided to the customer for reference. (For TEER measurements, MatTek has successfully used the WPI Endohm System.)
- Sterility: All media used throughout the production process is checked for sterility. Maintenance medium is incubated with and without antibiotics for 1 week and checked for sterility. The agarose gel from the 24-well plate used for shipping is also incubated for 1 week and checked for any sign of contamination.
- Screening for pathogens: All cells are screened and are negative for HIV, hepatitis B and hepatitis C using PCR. However, no known test method can offer complete assurance that the cells are pathogen free. Thus, these products and all human derived products should be handled at BSL-2 levels (biosafety level 2) or higher as recommended in the CDC-NIH manual, “Biosafety in microbiological and biomedical laboratories,” 1998. For further assistance, please contact your site Safety Officer or MatTek technical service.
- Notification of lot failure: If a tissue lot fails our QC or sterility testing, the customer will be notified and the tissues will be replaced without charge when appropriate. Because some of our QC and sterility testing is done post-shipment, notification will be made as soon as possible. (Under normal circumstances, ET-50 failures will be notified by Thursday, 5 p.m.; sterility failures will be notified within 8 days of shipment.)