EpiAirway™ Technical Specifications

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I. Ordering

1. 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.
2. ALL Tissue Orders: Please allow 3-4 weeks from order date to delivery of tissues. Please contact MatTek Customer Service for additional details.

II. Cells

1. Type: Normal human tracheal/bronchial epithelial (NHBE) cells.
2. Genetic make-up: Single donor.
3. Derived from: Non-smoker free of respiratory tract disease.
4. Screened for: HIV, Hepatitis-B, Hepatitis-C, mycoplasma, bacteria, yeast, fungi.
5. NOTE: EpiAirway tissues produced from ASTHMATIC, SMOKER and COPD donor cells are also available - please call the EpiAirway Lead Scientist for technical details, pricing and availability.

III. Medium

1. Base medium: Dulbecco's Modified Eagle's Medium (DMEM).
2. Growth factors/hormones: Epidermal growth factor and other proprietary factors.
3. Serum: None.
4. Antibiotics: Gentamicin 5 µg/ml (10% of normal gentamicin level).
5. Anti-fungal agent: Amphotericin B 0.25 µg/ml.
6. pH Indicator: Phenol red.
7. Other additives: proprietary.
8. Alternatives: Phenol red-free, antibiotic-free, anti-fungal-free medium and tissue are available. Agents are removed 3 days prior to shipment.
9. Assay medium: AIR-100-ASY.
10. Maintenance medium: AIR-100-MM.

IV. Tissue

1. Kits: Standard EpiAirway kit (AIR-100) consists of 24 tissues. EpiAirway Snapwell kit (AIR-100-SNP) consists of 24 tissues. AIR-606 kit consists of 6 tissues. (Tissue "kits" contain tissues, a small amount of culture medium, and plasticware; contact MatTek for specific kit contents.)
   
   
2. Substrate:
   AIR-100: Chemically modified, 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 = 0.9 cm. Surface area = 0.6 cm²).
   
   AIR-100-SNP: Costar Snapwell™ single well tissue culture plate inserts are used. (Pore size = 0.4 µm, Inner Diameter = 1.2 cm. Surface area = 1.0 cm²).
   Note: Snapwells fit in VITROCELL for 24 mm Transwells (need adaptor for Costar SnapWells - available upon request from Vitrocell).
   
   AIR-100-PC12: Transwell single well tissue culture plate inserts, polycarbonate membrane (Pore size = 0.4 µm, Inner Diameter = 1.2 cm. Surface area = 1.0 cm²).
   Note: Transwells fit in VITROCELL for 12 mm Transwells.
   
   AIR-606: Millipore Millicell™ CM single well tissue culture plate inserts. (Pore size = 0.4 µm, Inner Diameter = 2.2 cm. Surface area = 4.2 cm²).
   
   
3. Culture: At air liquid interface.
   
   
4. Histology: 3-4 cell layers, pseudo-stratified morphology. Displays mucociliary phenotype.
   
   
5. 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.
   
   
6. Shipment: At 4°C on medium-supplemented, agarose gels.
   
   
7. Shipment day: Monday.
   
   
8. Delivery: Tuesday morning via FedEx priority service (US). Outside US: Tuesday-Thursday depending on location.
   
   
9. Shelf life: Including time in transit, tissues may be stored at 4°C for up to 3 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 equilibration at 37°C.
   
   
10. Length of experiments: Tissue cultures can be continued for ONE (1) MONTH or more with good retention of normal morphology. Tissues must be fed every other day with 5.0 ml of maintenance medium (AIR-100-MM) Cell culture inserts are placed atop washers (EPI-WSHR) or culture stands (AIR-STND (same as MEL-STND; see photo below)) in 6-well plates to allow us of 5.0 ml.
    
    
11. Alternative Tissues/Forms:
    
    Alternative EpiAirway Tissue Types:
    
    EpiAirway tissues produced from ASTHMATIC, SMOKER and COPD donor cells are also available - please call the EpiAirway Lead Scientist for technical details, pricing and availability.

Asthmatic, Smoker, COPD EpiAirway Tissues

In addition to Standard EpiAirway Tissues produced from normal tracheal/bronchial epithelial cells of NON-SMOKING donors free of respiratory tract disease, EpiAirway tissues produced from ASTHMATIC, SMOKER and COPD donor cells are now available.

MEL-STND Accessory For Extended Culture Periods: Shown: 6-well plate with cell culture insert atop a culture stand (Part #MEL-STND). For experiments extending beyond 24 hours, tissues need to be fed with 5.0 ml every other day. Use of the culture stand allows feeding with 5.0 ml from the basolateral side of the tissue. Use of culture stands is necessary in order to maintain normal tissue morphology.

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Standard EpiAirway Tissue - Alternative Formats:

AIR-112: Same as AIR-100 except 12 tissues instead of 24.

AIR-100-SNP: Same as AIR-100 except tissues are cultured in Snapwell inserts which are compatible with permeability devices designed to handle Snapwell inserts.

AIR-100-PC12: Same as AIR-100 except tissues are grown on Transwell Inserts with polycarbonate membrane. For use in VITROCELL for 12 mm Transwells.

AIR-112-PC12: Same as AIR-100-PC12 except 12 tissues instead of 24.

AIR-124-HTS: Same as AIR-100 except 24 tissues are cultured in one 24-well Plate--for High Content Screening applications.

AIR-196-HTS: Same as AIR-100 except 96 tissues are cultured in one 96 Microwell Plate--for High Content Screening applications.

AIR-606: Same as AIR-100 except 6 tissues per kit, each tissue cultured in a larger diameter cell culture insert (22 mm diameter).

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EpiAirway-FT Full Thickness Tissues:

AFT-100: Highly differentiated, full-thickness human airway model composed of fibroblast-containing collagen matrix and airway epithelium cells.

AFT-112: Same as AFT-100 except 12 tissues instead of 24.

AFT-100-PC12: Same as AFT-100 except tissues are grown on Transwell Inserts with polycarbonate membrane. For use in VITROCELL for 12 mm Transwells.

AFT-112-PC12: Same as AFT-100-PC12 except 12 tissues instead of 24.

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EpiAirway RNA, Protein Lysate & Mucin

AIR-100-MUC: EpiAirway Mucin prepared from AIR-100, 1.0 ml minimum quantity (1 mg/ml). Inquire about product specifics.

AIR-100-PROT: EpiAirway Total Protein Lysate prepared from AIR-100, 100 ug. Inquire about specific treatments (PMA, UV, etc.).

AIR-100-RNA: Purified total RNA from control (untreated) or treated EpiAirway tissues. Each lot of RNA is characterized and provided with a quality control spec sheet including histology and Triton ET-50 of the specific tissue lots, agarose gel electrophoresis of total RNA, RNA concentration, 260/280 ratio, G3PDH RT-PCR and TAQ-only PCR amplifications. Control (untreated), cytokine treated (individually with IL-4, IL-13, TNF-α, as well as IL-4 + TNF-α, and IL-13 + TNF-α), and phorbol ester treated EpiAirway RNA's are currently available, 10 ug per vial.

V. Quality Control and Sterility

1. Visual inspection: All tissues are visually inspected and if physical imperfections are noted, tissues are rejected for shipment.
2. End-use testing: Transepithelial electrical resistance (TEER) of each lot is measured prior to shipment (2001 Calendar Year Average: 584 +139 Ohm • cm²). TEER in customers' lab may differ slightly from the MatTek results. A summary of these results will be provided to the customer for reference. (For TEER measurements, MatTek has successfully used the WPI Endohm System.)
3. 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.
4. 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.
5. 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).

EpiAirway Shipment and Dimension Information

Part Number Ship Day Tissue  Diameter Culture Area Number of Cultures AIR-100 Monday 8 mm 0.6 cm2 24 AIR-100-SNAP Monday 10 mm 1.0 cm2 12 AIR-100-ASTHMA Monday 8 mm 0.6 cm2 24 AIR-606 Monday 22.5 mm 4.2 cm2 6 AIR-100-HTS-24 Monday 0.3 mm 0.3 cm2 24