ImmunoPure^™ HD Natural Killer Cells CD56+

• 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 CD56+ Natural Killer (NK) cells are a type of cytotoxic lymphocyte that constitutes a major component of the innate immune system. NK cells play a major role in the rejection of tumors and cells infected by viruses. They kill cells by releasing small cytoplasmic granules of proteins called perforin and granzyme, which cause the target cell to die by apoptosis.

The ImmunoPure™ CD56+ Natural Killer (NK) 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™ CD56+ Natural Killer (NK) Cells
PBMC007A - 5.0 x 10⁶ cells - $800

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, and 10 IU/ml human IL-2.

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. 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 moistened with 70% alcohol; avoid wiping away the labeling.
5. Within about 30 sec., slowly add one milliliter of medium (containing serum) to the thawed cells.
6. 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.
7. Centrifuge for 5 min at 400 x g.
8. Aspirate or decant the supernatant and gently resuspend the cell pellet in 10 mL of medium.
9. Remove an aliquot for cell count and proceed with experimental manipulations.
10. Culture the cells in RPMI1640 supplemented with 10% fetal calf serum (heat inactivated), 2mM L-Glutamine, 100 IU/ml penicillin, 100 mg/ml streptomycin, and 10 IU/ml human IL- 2.

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.

C. References

1. Genetic modification of primary natural killer cells overcomes inhibitory signals and induces specific killing of leukemic cells. Chihaya Imai, Shotaro Iwamoto and Dario Campana. Blood; 2005 106: 376-383.