BioWhittaker X-VIVO media systems

Lonza’s R&D efforts and collaboration with many clinical trials have provided Lonza with the tools and expertise to support the developments in adoptive immunotherapy, cancer therapy, genetic therapy and other cellular therapies.

Lonza’s philosophy for the development of serum-free formulations for use in cellular therapy is to provide a nutritionally complete and balanced environment for the cells. We have not included any exogenous growth factors, artificial stimulators of cellular proliferation or undefined supplements.

The products are devoid of any protein-kinase C stimulators and are suitable for the investigation of second messenger systems in the activation of human and murine lymphocytes. The formulations are complete and contain pharmaceutical grade human albumin, recombinant human insulin and pasteurized human transferrin.

All X-VIVO medium products are manufactured under current GMPs and are listed with the FDA in a product Master File. Permission to cross-reference the Master File may be obtained by contacting the Product Manager.

X-VIVO 10 medium

The X-VIVO 10 medium formulation is designed to support the generation of Lymphokine Activated Killer (LAK) cells in a serum-free environment. The original protocols involved the incubation of patient or normal donor Peripheral Blood Lymphocytes (PBL) at 1.0-3.0 x 106 cells/mL for a period of 3 days in the presence of 1,000 Cetus units of rIL-2/mL. Optimal LAK cell generation is achieved when peripheral blood lymphocytes are incubated for three to ten days at a density of 1.0-6.0 x106 cells/mL in the presence of 100-1000 Cetus units of rIL-2. X-VIVO 10 medium is available as a 1X liquid in several convenient configurations.

X-VIVO 15 medium

X-VIVO 15 medium is similar in composition to X-VIVO 10 medium and has been optimized for the proliferation of Tumor Infiltrating Lymphocytes (TIL) under serum-free conditions. The formulation supports the proliferation of purified CD3+ cells isolated from peripheral blood and human tumors. X-VIVO 15 medium has also been used to support the growth of human monocytes, macrophage cells and cell lines, PBL, granulocytes and natural killer (NK) cells. In addition, X-VIVO 15 medium provides a serum-free environment for the expansion of HUT-78 and related human lymphocytic cell lines.

X-VIVO 20 medium

X-VIVO 20 medium was developed and optimized to support the generation of LAK cells from monocyte-depleted PBL at high density. Initial cell densities between 2.0-3.0 x 107 cells/mL were successfully used to generate LAK cells. X-VIVO medium 20 may also be used as a growth medium for PBL and TIL.

Other applications of X-VIVO products include:

• Proliferation of PBL
• Proliferation of TIL
• Cryopreservation and transplantation of organs
• Cultivation of human monocytes and macrophages
• Cultivation of stem cells
• Cultivation of dendritic cells

Partial list of cell types grown with X-VIVO medium

• Human and murine Lymphokine Activated Killer (LAK) cells
• Peripheral Blood Lymphocytes
• Human and murine Tumor Infiltrating Lymphocytes (TIL)
• Human T cells
• Human and murine macrophages
• Human and murine hematopoietic stem cells (colony formation and proliferation)
• Cryopreservation of human tissue
• Proliferation of draining lymph node lymphocytes

X-VIVO Media References

X-VIVO 10 medium

1. Cavazzana-Calvo, M. and others. “Gene Therapy of Human Severe Combined Immunodeficiency (SCID) -X1 Disease.” Science 288 (April 2000): 669-672

2. Pawelec, G. and others. “T Cell Immunosenescence in Vitro and in Vivo.” Exp Gerontol 34, no. 3 (June 1999): 419-29.

3. Pawelec, G. and others. “Extrathymic T Cell Differentiation in Vitro From Human CD34+ Stem Cells.” J Leukoc Biol 64, no. 6 (December 1998): 733-9.

4. Weich, N. S. and others. “Recombinant human interleukin-11 directly promotes megakaryocytopoiesis in vitro.” Blood 90, no. 10 (November 1997): 3893-902.

5. Williams, Stephanie F. and others. “Selection and Expansion of Peripheral Blood CD34+ Cells in Autologous Stem Cell Transplantation for Breast Cancer.” Blood 87 (1996): 1687-91.

6. Sandstrom, Craig E. and others. “Comparison of Whole Serum-Deprived Media for Ex Vivo Expansion of Hematopoietic Progenitor Cells From Cord Blood and Mobilized Peripheral Blood Mononuclear Cells.” Journal of Hematotherapy 5 (1996): 461-73.

7. Zimmerman, Todd M. and others. “Clinical Use of Selected and Expanded Peripheral Blood CD34+ Cells: A Preliminary Report of Feasibility and Safety.” Journal of Hematotherapy 4 (1995): 527-29.

8. Purdy, Malcolm H. and others. “Large Volume Ex Vivo Expansion of CD34-Positive Hematopoietic Progenitor Cells for Transplantation.” Journal of Hematotherapy 4 (1995): 515-25

9. Abrahamsen, T. G. and others. “Stimulatory Effect of Counterflow Centrifugal Elutriation in Large-Scale Separation of Peripheral Blood Monocytes Can Be Reversed by Storing the Cells at 37 Degrees C.” J. Clin. Apheresis 6 (1991): 48-53.

10. Streck, Richard J. and others. “Lysis of Autologous Human Macrophages by Lymphokine-Activated Killer Cells: Interaction of Effector Cell and Target Cell Conjugates Analyzed by Scanning Electron Microscopy.” Journal of Leukocyte Biology 48 (1990): 237-46.

11. Helinski, Ernest H. and others. “Tumor- Cytolytic Human Macrophages Cultured As Nonadherent Cells: Potential for the Adoptive Immunotherapy of Cancer.” Cancer Detection and Prevention 14, no. 4 (1990): 471-81.

X-VIVO 15 medium

1. Kugler, Alexander and others. “Regression of Human Metastatic Renal Cell Carcinoma After Vaccination With Tumor Cell - Dendritic Cell Hybrids.” Nature Medicine 6, no. 3 (March 2000): 332-36.

2. Yang, Shiaolan and others. “Generation of Retroviral Vector for Clinical Studies Using Transient Transfection.” Human Gene Therapy 10 (January 1999): 123-32.

3. Chen, Bing-guan and others. “The Role of Tumor Necrosis Factor Alpha in Modulating the Quantity of Peripheral Blood-Derived, Cytokine-Driven Human Dendritic Cells and Its Role in Enhancing the Quality of Dendritic Cell Function in Presenting Soluble Antigens to CD4+ T Cells In Vitro.” Blood 91 , no. 12 (1998): 4652-61.

4. Jonuleit, H. and others. “Pro-Inflammatory Cytokines and Prostaglandins Induce Maturation of Potent Immunostimulatory Dendritic Cell Under Fetal Calf Serum-Free Conditions.” Eur J Immunol 27, no. 12 (December 1997): 3135-42.

5. Steinbrink, K. and others. “Induction of Tolerance by IL-10 Treated Dendritic Cells.” J Immuno 159, no. 10 (November 1997): 4772-80.

6. Jonuleit, H. and others. “Induction of IL-15 Messenger RNA and Protein in Human Blood-Derived Dendritic Cells: a Role for IL-15 in Attraction of T Cells.” J Immunol 158, no. 6 (March 1997): 2610-5.

7. Strobl, Herbert and others. “Flt3 Ligand in Cooperation with Transforming Growth Factor - Beta-1 Potentiates in Vitro Development of Langerhans-Type Dendritic Cells and Allows Single-Cell Dendritic Cell Cluster Formation Under Serum-Free Conditions.” Blood 90, no. 4 (1997): 1425-34.

X-VIVO 20 medium

1. Luft, T. and others. “Type I IFNs Enhance the Terminal Differentiation of Dendritic Cells.” J Immunol 161, no. 4 (August 1998): 1947-53.

2. Luft, T. and others. “A Serum-Free Culture Model for Studying the Differentiation of Human Dendritic Cells From Adult CD34+ Progenitor Cells.”Exp Hematol 26, no. 6 (June 1998): 489-500.

3. Jonuleit, H. and others. “Pro-Inflammatory Cytokines and Prostaglandins Induce Maturation of Potent Immunostimulatory Dendritic Cell Under Fetal Calf Serum-Free Conditions.” Eur J Immunol 27, no. 12 (December 1997): 3135-42.

 

Storage: 2 to 8°C

 

Product Use Statement:

THESE PRODUCTS ARE FOR RESEARCH USE ONLY. Not approved for human or veterinary use, for application to humans or animals, or for use in clinical or in vitro procedures.