ImmunoPure^™ HD T Cells CD8+

• 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 CD8+ Cytotoxic T Cells are a sub-group of T lymphocytes capable of inducing the death of infected -with viruses or other pathogenssomatic or tumor cells. Most CD8+ Cytotoxic T cells express T-cell receptors that can recognize a specific antigenic peptide bound to Class I MHC molecules (present on all nucleated cells) and a glycoprotein called CD8. The MHC molecule keeps the Cytotoxic T cell and the target cell bound closely together during antigen-specific activation. CD8+ T cells are recognized as Cytotoxic T cells once they become activated and are generally classified as having a pre-defined cytotoxic role within the immune system.

ImmunoPure™ HD T Cells CD8+ 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™ HD T Cells CD8+
PBMC004A - 5.0 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

RPMI 1640 medium + 10% (v/v) FBS, 2 mM glutamine, 1% (v/v) nonessential amino acids, 1% (v/v) sodium pyruvate, 50 U/ml penicillin, 50 mg/ml streptomycin, and 50 mg/ml kanamycin.

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.
11. Culture the cells in RPMI 1640 medium supplemented with 10% (v/v) FBS, 2 mM glutamine, 1% (v/v) nonessential amino acids, 1% (v/v) sodium pyruvate, 50 U/ml penicillin, 50 mg/ml streptomycin, and 50 mg/ml kanamycin at a density of 1-2 million cells/ml.

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.