ImmunoPure^™ HD Dendritic Cells

• Proprietary reagents ensure high survival and recovery rates
• Analyzed and characterized by Flow cytometry
• Each lot tested to be free from infectious agents and contaminants

Normal Human Dendritic cells (DCs) are immune cells that form part of the mammalian immune system. Their main function is to process antigen material and present it on the surface to other cells of the immune system, thus functioning as antigen-presenting cells. They act as messengers between the innate and adaptive immunity. Dendritic cells are present in small quantities in tissues that are in contact with the external environment, mainly the skin and the inner lining of the nose, lungs, stomach and intestines. They can also be found in an immature state in the blood. Once activated, they migrate to the lymphoid node where they interact with T cells and B cells to initiate and shape the adaptive immune response.

The ImmunoPure™ Dendritic Cells are isolated from fresh human adult peripheral blood of healthy donors at IRB-approved and FDAlicensed blood banks, using apheresis and immunomagnetic cell separation techniques.

Because of their importance in multiple life science research areas, Genlantis is pleased to introduce the ImmunoPure NP line of peripheral blood cells. They consist of a variety of immune cells derived from the peripheral blood of normal adults, thereby qualifying them for a diverse number of cell-based experiments.

ImmunoPure™ Dendritic Cells
PBMC008A - 0.5 x 10⁶ cells - $700

Figure 1. Figure 1: Third Party Fresh vs Thawed Viability Test

ImmunoPure NP Normal Human Cells are isolated from the peripheral blood (buffy coats) of healthy human adult donors performed at IRB-approved and FDA-licensed blood banks across the United States. The lymphocyte and monocyte fractions are isolated from whole blood using apheresis, a technology in which the blood of a donor or patient is passed through an apparatus that separates out these fractions, and returns the remainder to the donor’s circulation. After collection, the cells are treated with Ficoll® in order to deplete granulocytes, and red blood cells. These PBMCs are immediately cryopreserved by decreasing the temperature by 1 degree Celsius/minute down to -170ºC and using proprietary reagents to ensure high survival and recovery rates. ImmunoPure NP cells are high quality products and research tools that represent a diverse and heterogeneous population of donors. Genlantis ImmunoPure NP cells are backed by Genlantis’ customer satisfaction guarantee*. All cell batches are rigorously tested to be free from infectious agents and contaminants such as HIV-1, HBV, HCV, syphilis, bacterial, fungus, and mycoplasma. Our cells are routinely analyzed and characterized by Flow cytometry, using cell viability, cell purity, cell count, and CD markers (like CD34 and CD45) as indicators; this information is provided with each shipment on a lot-specific certificate of analysis. Recent independent third party testing have confirmed the high viability of the ImmunoPure NP cells (Figure 1).

ImmunoPure NP cells are shipped on dry ice. It is highly recommended that the cells are used as soon as possible after receipt in order to obtain the highest viability possible. For short term (3 weeks or less) cells should be stored at -80ºC. For longer term storage, cells should be kept in liquid nitrogen (-170ºC). Because of the nature of these cells, users should note that viability cannot be guaranteed after storage at -170ºC storage or beyond one week at -80ºC.

A. General Medium Requirement

RPMI1640 + 10% fetal calf serum (heat inactivated), 2mM LGlutamine, 100 IU/ml penicillin, 100 mg/ml streptomycin.

B. Thawing and Culturing Cells

1. Prepare a 37ºC water bath to temperature.
2. Keep all samples frozen until the bath is ready.
3. Place the vials into the water bath, being careful not to submerge below the junction between the lid and the vial.
4. Place vial of cells in 37°C water bath and agitate until thawed. It is important to thaw the cells quickly; do NOT allow thawed cells to remain in freezing media any longer than necessary.
5. When only a small amount of ice remains, remove the vials and dry with a lab tissue. Clean the top of the vial with a lab tissue wetted with 70% alcohol; avoid wiping away the labeling.
6. Within about 30 sec., slowly add one milliliter of medium (containing serum) to the thawed cells.
7. Slowly add thawed cells to 8 mL of medium containing serum. Invert tube 2 or 3 times to mix or mix gently by pipetting up and down several times.
8. Centrifuge for 5 min. at 400 x g.
9. Aspirate or decant the supernatant and gently resuspend the cell pellet in 10 mL of medium.
10. Remove an aliquot for cell count and proceed with experimental manipulations.
NOTE: The cell suspension may form clumps after standing at room temperature. This can be avoided by preparing and using the cells promptly or by adding DNase to the suspension at a final concentration of 10 units per ml.
11. Culture the cells in RPMI 1640 supplemented with 10% heat inactivated fetal calf serum (FCS), 2 mM L-glutamine, 100 IU/ml penicillin and 100 mg/ml streptomycin at a density of 1-2 million cells/ml.
NOTE: Dendritic cells do not proliferate. They will survive in culture for 1 week or more. If cells are to be maintained longer than overnight, addition of 500 U/ml GM-CSF and 500 U/ml IL4 is recommended to ensure the DC phenotype is maintained.