Serum-free T-Cell Expansion Medium has been developed to promote the optimal expansion of T-Cells of human origin. This medium supports high viable cell densities. The elimination of serum reduces performance variability in the medium and eliminates safety risks associated with possible adventitious agents in serum. The set of Serum-free Medium  consists of two parts, Culture Medium (Nobimpex 003)  and Growth-Promoting Solution (Nobimpex 004). T-Cell Growth-Promoting Solution(Nobimpex 004) for the expansion of T-cells during the first five days, and Serum-free T-Cell Culture Medium(Nobimpex 003) be used in another days.

More Safety:

The Serum-free medium set are proprietary formulation. And both the medium do not contain antibiotics or cytokines. Human serum albumin and human transferrin are the only human origin materials and are non-reactive (donor level) for anti-HIV 1 & 2, anti-HCV and HBsAg. Handle as if potentially infectious.

More stringent quality and safety control.

Preparation of complete media:

The medium set is supplied as a sterile (1X) liquid and must be supplemented with stimulatory antibodies, cytokines and/or antibiotics, if desired. Serum-free T-Cell Expansion Medium(Nobimpex 003) must also be supplemented with L-glutamine, add 20ml of 200mM L-glutamine solution or 0.584 g powder (irradiated) per liter of medium.

We recommend that Nobimpex 004 medium be used in two times(25 ml each) during the first five days Expansion of T cells , and Nobimpex 003 medium be added as well in another days .

 Procedure:

1. Add certain amount of stimulatory antibodies and cytokines in 25 ml Nobimpex 004 medium.

2.Prepare either fresh or frozen PBMNCs (peripheral blood mononuclear cells) as directed by the supplier or in accordance with established protocols.

3. Count cells using a hemacytometer.

4.Transfer the proper number of cells to the desired culture vessel containing medium supplemented with cytokines and stimulatory antibodies (and antibiotics if desired).

5. Place the culture vessel in a humidified in cubator at 37°C and 5% CO2.

6. Day 3, add other 25ml the Nobimpex 004 medium supplemented with cytokines and stimulatory antibodies(and antibiotics if desired).

7. Day 5 and beyond, add Nobimpex 003 medium which also supplemented with cytokines and stimulatory antibodies (and antibiotics if desired) with optimal amount.

Product Profile:

Nobimpex’s Serum-freeT-Cell Expansion Medium set(Nobimpex 003&004) demonstrated rigorous expansion of T-cells from PBMNCs. This product was compared with several other commercially available serum-free expansion media for their ability to expand T-cells in T75 culture flasks. For these small-scale experiments, 200,000 PBMNCs/ml were incubated for up to 7 days in Serum-free medium or other commercial product containing 100 IU/ml IL-2 and anti-CD3 (OKT3, 20ng/ml) antibody.

The expanded T-Cell population from these experiment was then used in 51 chromium release assays to test for functional cytolytic potential. Briefly, target cells (K562, a human chronic myelogenous leukemia cell line), were labeled with 51 Chromium and linked to anti-CD3 (OKT3) via Fc receptor. When mixed with effector orcytolytic T-cells, the target cells under go apoptosis or lysis and release 51 Chromium. The amount of 51 chromium release into the supernatant is proportional to the number of targets killed and the number of functional cytolytic T-Cells.