AIM-V® Medium CTS™
Therapeutic Grade rum free cell expansion medium
GIBCO® AIM-V Medium CTS™ (Therapeutic Grade) is the first commercially available defined, rum-free formulation for proliferation and/or manipulation of T-cells and dendritic cells and manufactured in compliance with cGMP. AIM-V Medium CTS™ is an FDA 510(k) cleared device which is intended for human ex-vivo tissue & cell culture processing applications.
Description Cat. No. Size
AIM-V® Medium CTS™, Liquid 0870112DK 1000mL AIM-V® Medium CTS™, Liquid 0870112BK 10L
(Bag)
Intended
U
For human ex-vivo tissue & cell culture processing applications. CAUTION: When ud as a medical device, Federal Law restricts this device to sale by or on the order of a physician. Storage
Store medium at 2 to 8°C. Protect from light.
Shelf Life
14 months
Culture Procedure:
The procedure below rves as a general guideline for static T- cell and dendritic cell culture, regardless of vesl. For high-density culture in bioreactors, optimal procedures should be determined empirically by the investigator.
T Cells Culture:
1. Prepare fresh peripheral blood mononuclear cells (PBMCs)
or rapidly thaw (< 1 minute) frozen vials of PBMCs cells in a 37°C water bath according to standard PBMC thawing protocols.
2. Wash cells with DPBS CTS™ without calcium and
magnesium (Cat. No A12856), with 2-5% heat-inactivated human pooled Type AB rum according to the applications, if desired or required.
3. Count cells using either electronic (i.e. Coulter Counter, Vi-
Cell) or manual (i.e. hemocytometer) methods.
4. Centrifuge cells and remove wash buffer.
5. Resuspend PBMC at roughly 0.5-1x106 CD3+ T cells/mL in
medium supplemented with cytokines (e.g. IL-2), if ud at culture initiation. Transfer the desired number of cells to the desired tissue culture vesl. A variety of protocols may be ud for activating T-cells for subquent expansion, including adding stimulatory antibodies or antigen prenting cells. Similarly, for either small or the large scale T-cell expansion, cells can be isolated, activated an
d expanded with Dynabeads® ClinExVivo™ CD3/CD28 or Dynabeads® CD3/CD28 CTS TM(Cat. No. 402-03D) according to instructions in the product inrt. 6. Incubate the culture vesl at 37°C in a humidified
atmosphere with 5% CO2. Feed and maintain cells at desired concentrations while cells are in log pha growth.
To maintain log pha growth, it may be preferable to split cells to achieve a density of 0.5-1x106cells/mL whenever cell density gets above 1x106cells/mL (e.g. 2x106cells/mL, split 1:4 to continue culture at 0.5x106cells/mL). For optimal gas exchange in static plate cultures it is recommended that medium depth not exceed 1 to 1.2cm.
Monocyte Derived Dendritic Cell Culture:
1. Prepare fresh peripheral blood mononuclear cells (PBMCs).
2. Plate PBMC in culture flask with 25 mL RPMI 1640 (Cat.
No 72400) or AIM-V® Medium CTS™ (Therapeutic Grade).
3. Incubate for 2 to 3 hours at 36 to 38
˚C in a humidified atmosphere of 5% CO2 in air.
4. Discard medium containing non-adherent cells.
5. Wash the adherent cells (mainly CD14+ monocytes) three
times with DPBS without calcium and magnesium (Cat. No A12856).
6. Add medium containing 50 to 100 ng/mL recombinant
human IL-4 (Cat. No. CTP0043 1mg or Cat. No. CTP0041 100ug) and 50 ng/mL recombinant human GM-CSF (Cat.
No. CTP2011 100ug or Cat. No. CTP2013 1mg). Cell density should be between 1 to 3x105 cells/mL.
7. Incubate cells at 36 to 38˚C in a humidified atmosphere of
5% CO2 in air for 5 days. It is recommended to replace medium once after 3 days with fresh medium containing IL-
4 and GM-CSF. Save all non-adherent or looly adherent
cells by centrifuging the removed culture medium 10 minutes at 200xg and adding the pellet to the fresh culture medium.
8. After 6 days, the looly adherent or non-adherent cells
should display typical dendritic cell morphology and surface markers (CD1a, CD80, CD86, and HLA-DR).
9. The maturation of dendritic cells is induced by the addition
of either 1 µg/mL LPS or 50µl/mL TNF-α (cat. No. CTP3013 1mg or Cat. No. CTP3011 100ug) to the medium.
Note: Alternatively to plastic adherence, monocytes can also be isolated by magnetic paration.
Related Products
Dulbecco's Phosphate Buffered Saline CTS™ (DPBS) without calcium, magnesium (1X), liquid (A12856) L-Glutamine-200mM (100X), liquid (25030)
Dynabeads ClinExVivo™ CD3/CD28 ®
or Dynabeads CD3/CD28 ®
CTS TM (402-03D) DynaMag™ CTS™ (121-02)
IL-2 CTS™ REC HU (CTP0021 100ug or CTP0023 1mg) IL-7 CTS™ REC HU (CTP0071 100ug or CTP0073 1mg) IL-4 CTS™ REC HU (CTP0041 100ug or CTP0043 1mg) GM-CSF CTS™ REC HU (CTP2011 100ug or CTP2013 1mg) TNF-α CTS™ (CTP3011 100ug or CTP3013 1mg)
Technical Support
For additional product and technical information, such as Material Safety Data Sheets (MSDS), Certificate of Analysis,
etc, plea visit our website at /celltherapysupport/. For further assistance, plea email our Technical Support team at
The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners
References
1.
Rebecca J et al., (2010) Natural exposure to cutaneous anthrax gives long lasting T cell immunity encompassing infection-specific Epitopes. J. Immunol., 184: 3814 – 3821
2.
Fabricius D et al., (2010) Prostaglandin E2 inhibits IFN-α cretion and Th1 costimulation by human plasmacytoid dendritic cells via E-prostanoid 2 and E-prostanoid 4 receptor engagement. J. Immunol., 184: 677 – 684
3.
Nesbit L et al., (2010) Polyfunctional T Lymphocytes Are in the Peripheral Blood of Donors Naturally Immune to Coccidioidomycosis and Are Not Induced by Dendritic Cells . Infect. Immun., 78: 309 - 315
4. Jahrsdorfer B et al., (2010) Granzyme B produced by human plasmacytoid dendritic cells suppress T-cell expansion . Blood, 115: 1156 – 1165
5.
Csillag A et al., (2010) Pollen-Induced Oxidative Stress Influences Both Innate and Adaptive Immune Respons via Altering Dendritic Cell Functions. J. Immunol., 184: 2377 – 2385
6.
Cornberg M et al., (2010) CD8 T Cell Cross-Reactivity Networks Mediate Heterologous Immunity in Human EBV and Murine Vaccinia Virus Infections. J. Immunol., 184: 2825 - 2838.
7.
Bellone S et al., (2009) Human Papillomavirus Type 16 (HPV-16) Virus-Like Particle L1-Specific CD8 Cytotoxic T Lymphocytes (CTLs) Are Equally Effective as E7-Specific CD8 CTLs in Killing Autologous HPV-16-Positive Tumor Cells in Cervical Cancer Patients: Implications for L1 Dendritic Cell-Bad Therapeutic Vaccines ++. J. Virol., 83: 6779 - 6789
8. Sato K et al., (2009) Impact of culture medium on the expansion of T cells for immunotherapy. Cytotherapy 11: 4-11
9.
Liu ZW et al., (2009) A CD26-Controlled Cell Surface Cascade for Regulation of T Cell Motility and Chemokine Signals . J. Immunol ., 183: 3616 - 3624.
10. Megyeri M et al., (2009) Complement Protea MASP-1 Activates Human
Endothelial Cells: PAR4 Activation Is a Link between Complement and Endothelial Function. J. Immunol., 183: 3409 - 3416. 11.
Manfred L et al., (2005) Functional characterization of monocyte-derived dendritic cells generated under rum free culture conditions. Immunology letters 99: 209-216
12. Nagorn D et al., (2003) Biad epitope lection by recombinant vaccinia-virus (rVV)-infected mature or immature dendritic cells. Gene Therapy 10: 1754-1765 13. Lotem M et al., (2006) Prentation of tumor antigens by dendritic cells genetically modified with viral and nonviral vectors. J immunotherapy 29: 616-627
14.
Dietze B et al. (2008) An improved method to generate equine dendritic cells from peripheral blood mononuclear cells: divergent maturation programs by IL-4 and LPS. Immunobiology 213:751–758.
15.
Meehan KR et al. (2008) Development of a clinical model for ex vivo expansion of multiple populations of effector cells for adoptive cellular therapy. Cytotherapy 10: 30–37.
16. Ye Z et al. (2006) Human dendritic cells engineered to express alpha tumor
necrosis factor maintain cellular maturation and T-cell stimulation capacity. Cancer Biother Radiopharm 21:613–622.
17. Choi BH et al. (2006) Optimization of the concentration of autologous rum for
generation of leukemic dendritic cells from acute myeloid leukemic cells for clinical immunotherapy. J Clin Apher 21:233–240. 18.
Imataki O et al. (2006) Efficient ex vivo expansion of alpha24+ NKT cells derived from G-CSF-mobilized blood cells. J Immunother 29:320–327.
19. Peng JC et al. (2005) Generation and maturation of dendritic cells for clinical
application under rum-free conditions. J Immunother 28:599–609. 20. Trickett AE et al. (2002) Ex vivo expansion of functional T lymphocytes from HIV infected individuals. J Immunol Methods 262:71–83.
21.
Carlens S et al. (2000) Ex vivo T lymphocyte expansion for retroviral transduction: influence of rum-free media on variations in cell expansion rates and lymphocyte subt distribution. Exp Hematol 28:1137–1146.
22.
Kambe N et al. (2000) An improved procedure for the development of human mast cells from disperd fetal liver cells in rum-free culture medium. J Immunol Methods 240:101–110.
23. Gerin PA et al. (1999) Production of retroviral vectors for gene therapy with the
human packaging cell line FLYRD18. Biotechnol Prog 15:941–948. 24. Slunt JB et al. (1997) Human T-cell respons to Trichophyton tonsurans:
inhibition using the rum free medium Aim-V. Clin Exp Allergy 27:1184–1192. 25. Kreuzfelder E (1996) Asssment of peripheral blood mononuclear cell
proliferation by [2-3H]adenine uptake in the woodchuck model. Clin Immunol Immunopathol 78:223–227. 26.
Cauy AL (1994) A rum-free medium for human primary T lymphocyte culture. J Immunol Methods 175:115–121.
27. Freedman RS et al. (1994) Large-scale expansion in interleukin-2 of
tumorinfiltrating lymphocytes from patients with ovarian carcinoma for adoptive immunotherapy. J Immunol Methods 167:145–160. 28.
Nomura K et al. (1993) [Study of adoptive immunotherapy for metastatic renal cell carcinoma with lymphokine-activated killer (LAK) cells and interleukin-2. II. Clinical evaluation.] Nippon Hinyokika Gakkai Zasshi 84:831–840. Japane.
29.
Kaldjian EP et al. (1992) Enhancement of lymphocyte proliferation assays by u of rum-free medium. J Immunol Methods 147:189–195.
30. Hayakawa K et al. (1991) Study of tumor-infiltrating lymphocytes for adoptive
therapy of renal cell carcinoma (RCC) and metastatic melanoma: quential proliferation of cytotoxic natural killer and noncytotoxic T cells in RCC. J Immunother 10:313–325. 31.
McVicar DW et al. (1991) A comparison of rum-free media for the support of in vitro mitogen-induced blastogenic expansion of cytolytic lymphocytes. Cytotechnology 6:105–113.
32. Burg S et al. (1991) [Effect of different media on long-term cultivation of human
synovial macrophages.] Z Rheumatol 50:142–150. German. 33. Helinski EH et al. (1988) Long-term cultivation of functional human macrophages
in Teflon dishes with rum-free media. J Leukoc Biol 44:111–121. 34. Robyn S et al. (2007) RA8, A human anti-CD25 antibody against human Treg
cells. Hybridoma 26:119–130. 35.
Chena X et al. (2006) Induction of primary anti-HIV CD4 and CD8 T cell respons by dendritic cells transduced with lf-inactivating lentiviral vectors. Cell Immunol 243:10–18.
36. Grant R et al. (2008) CCL2 increas X4-tropic HIV-1 entry into resting CD4+ T
cells. J Biol Chem 283:30745–30753. 37.
Hagihara M et al. (2003) Incread frequency of CD3/8/56-positive umbilical cord blood T lymphocytes after allo-priming in vitro. Ann Hematol 82:166–170.
38. Wang Z et al. (2006) Application of rum-free culture medium for preparation of
A-NK cells. Cell Mol Immunol 3:391–395. 39. Morecki S et al. (1991) Retrovirus-mediated gene transfer into CD4+ and CD8+
human T cell subts derived from tumor-infiltrating lymphocytes and peripheral blood mononuclear cells. Cancer Immunol Immunother 32:342–352. 40.
Johann P et al. (2003) CD4 T cells guarantee optimal competitive fitness of CD8 memory T cells. Eur J Immunol 34:91–97.
June 2010
Form No. 5047
