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.

II. Cells

1. Type: Normal human dendritic/Langerhans cells. Cells are available ready-to-use (DC-100-RT) or cryopreserved (DC-100-CRY).
2. DC Subsets: DC-100 contains both plasmacytoid DC (pDC; CD123+, CD11c-) and myeloid DC (mDC; CD123-, CD11c+).
3. Genetic make-up: Single donor. Many donors are available (>15).
4. Generated from: CD34+ progenitor cells derived from umbilical cord blood.
5. Intended use: For in vitro research purposes only.

III. Medium

1. Base medium: RPMI-1640.
2. Serum: Fetal bovine serum.
3. Antibiotics: Penicillin/Streptomycin (Normal level).
4. Anti-fungal agent: None
5. pH Indicator: Phenol red.
6. Other additives: Cytokines and other factors (proprietary formulation).
7. Basal medium: The basal medium (DC-BAS-25, 25 ml or DC-BAS-250, 250 ml) is the basal medium used in the maintenance medium (DC-MM-25, DC-MM-50, or DC-MM-250; 25, 50 or 250 ml, respectively). The basal medium does not contain cytokines. Shelf life = 1 month. The DC-100-RT cells are shipped in a T25 flask containing DC-BAS. DC-BAS-25 is included with each vial of DC-100-CRY.
8. Maintenance medium: The maintenance medium (DC-MM-25, DC-MM-50 or DC-MM-250) contains a proprietary mixture of cytokines which are added to maintain the dendritic cell phenotype (see III. a. below). The maintenance medium is also available in 50 ml (DC-MM-50) aliquots. Shelf life = 2 weeks.
   Note: Culturing DC at a density of 250,000 cells/ml in DC-MM is recommended for optimal results.

IV. Dendritic/Langerhans Cells

1. DC-100: Both ready-to-use (DC-100-RT) and cryopreserved dendritic cells (DC-100-CRY) contain 3.0 - 3.5 million cells.
2. Culture: Suspension culture.
3. Phenotypic Markers and TLR's: Birbeck granules, CD1a, HLA-DR, CD11c, CD40, CD54, CD80, CD86, CD123, CD206, Langerin, TLR-1, TLR-2, TLR-3, TLR-4, TLR-6, TLR-7, TLR-9.
4. Lot Numbers: The lot number of the DC-100 cells identifies the date of shipment and donor. All cells within a specific lot were harvested, cryopreserved, and cultured together and are identical in regards to cells, medium, handling, culture conditions, etc.
5. Shipment: Cryopreserved DC (DC-100-CRY) are shipped in a cryovial on dry ice. Ready-to use DC are shipped in a T25 culture flask at ambient temperature in basal medium (DC-BAS).
6. Shipment day: For US: Every Monday, Tuesday, or Wednesday. For Europe: Every Monday or Tuesday.
7. Ordering: Orders must be placed a minimum of 24 hours prior to the desired shipment date.
8. Delivery: For US: Morning following shipment via FedEx priority service. Outside US: 1-3 days following shipment depending on location.
9. Shelf life: The shelf life of DC-100-CRY is unlimited provided that the cells arrive frozen and that they are placed in liquid nitrogen upon arrival. DC-100-RT are meant to be used/processed on the day of arrival.
10. Length of experiments: Cultures can be maintained for 10 - 15 days with good retention of normal dendritic cell characteristics and function. Cultures must be fed every other day with 1 ml DC-MM medium per 250,000 cells.
    
    Note: DC Maintenance medium (DC-MM-25, 25 ml) is supplied with each cryovial (DC-100-RT) or T25 flask (DC-100-RT) of cells. If the cells will be cultured for more than 2 days, additional maintenance medium (DC-MM-50 or DC-MM-250) will be needed. Discussion with a MatTek technical representative is strongly recommended prior to ordering due to the complex and proprietary nature of this product.
    
    
11. Alternative Cells/Forms:
    
    DCP-100: Plasmacytoid Dendritic Cells. Both ready-to-use (DCP-100-RT) and cryopreserved dendritic cells (DCP-100-CRY) contain 3.0 - 3.5 million cells.
    
    DC-100-RNA: Purified total RNA from control (untreated) or treated DC-100 Dendritic Cells. Each lot of RNA is provided with a quality control spec sheet including agarose gel electrophoresis of total RNA, RNA concentration, and 260/280 ratio. Control (untreated) Dendritic Cell RNAs (10 ug) are available from stock. Treated Dendritic Cell RNAs are available upon request.

V. Related Products

1. T-cells: T-cells which are autologous with the DC-100 are available upon request.

VI. Quality Control and Sterility

1. Characterization: Cells are characterized for expression of the surface markers CD1a (>40%), HLA-DR (>75%) prior to cryopreservation.
2. Sterility: All media used throughout the production process is checked for sterility. Maintenance medium is incubated with and without antibiotics for 2 weeks to check for sterility.
3. 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.