Accelerated production of antigen-specific T cells for preclinical and clinical applications using gas-permeable rapid expansion cultureware (G-Rex)

Juan F. Vera, Lara J. Brenner, Ulrike Gerdemann, Minhtran C. Ngo, Uluhan Sili, Hao Liu, John Wilson, Gianpietro Dotti, Helen E. Heslop, Ann M. Leen, Cliona M. Rooney

Research output: Contribution to journalArticle

86 Citations (Scopus)

Abstract

The clinical manufacture of antigen-specific cytotoxic T lymphocytes (CTLs) for adoptive immunotherapy is limited by the complexity and time required to produce large numbers with the desired function and specificity. The culture conditions required are rigorous, and in some cases only achieved in 2-cm 2 wells in which cell growth is limited by gas exchange, nutrients, and waste accumulation. Bioreactors developed to overcome these issues tend to be complex, expensive, and not always conducive to CTL growth. We observed that antigen-specific CTLs undergo 7 to 10 divisions poststimulation. However, the expected CTL numbers were achieved only in the first week of culture. By recreating the culture conditions present during this first week-low frequency of antigen-specific T cells and high frequency of feeder cells-we were able to increase CTL expansion to expected levels that could be sustained for several weeks without affecting phenotype or function. However, the number of 24-well plates needed was excessive and cultures required frequent media changes, increasing complexity and manufacturing costs. Therefore, we evaluated novel gas-permeable culture devices (G-Rex) with a silicone membrane at the base allowing gas exchange to occur uninhibited by the depth of the medium above. This system effectively supports the expansion of CTL and actually increases output by up to 20-fold while decreasing the required technician time. Importantly, this amplified cell expansion is not because of more cell divisions but because of reduced cell death. This bioprocess optimization increased T-cell output while decreasing the complexity and cost of CTL manufacture, making cell therapy more accessible.

Original languageEnglish (US)
Pages (from-to)305-315
Number of pages11
JournalJournal of Immunotherapy
Volume33
Issue number3
DOIs
StatePublished - Apr 1 2010
Externally publishedYes

Fingerprint

Cytotoxic T-Lymphocytes
Gases
T-Lymphocytes
Antigens
Feeder Cells
Adoptive Immunotherapy
Costs and Cost Analysis
Lymphocyte Count
Bioreactors
Silicones
Growth
Cell- and Tissue-Based Therapy
Cell Division
Cell Death
Phenotype
Food
Equipment and Supplies
Membranes

Keywords

  • Antigen-specific cytotoxic T lymphocytes
  • Immunotherapy
  • Large-scale expansion
  • Novel gas-permeable cultureware

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Pharmacology
  • Cancer Research

Cite this

Accelerated production of antigen-specific T cells for preclinical and clinical applications using gas-permeable rapid expansion cultureware (G-Rex). / Vera, Juan F.; Brenner, Lara J.; Gerdemann, Ulrike; Ngo, Minhtran C.; Sili, Uluhan; Liu, Hao; Wilson, John; Dotti, Gianpietro; Heslop, Helen E.; Leen, Ann M.; Rooney, Cliona M.

In: Journal of Immunotherapy, Vol. 33, No. 3, 01.04.2010, p. 305-315.

Research output: Contribution to journalArticle

Vera, JF, Brenner, LJ, Gerdemann, U, Ngo, MC, Sili, U, Liu, H, Wilson, J, Dotti, G, Heslop, HE, Leen, AM & Rooney, CM 2010, 'Accelerated production of antigen-specific T cells for preclinical and clinical applications using gas-permeable rapid expansion cultureware (G-Rex)', Journal of Immunotherapy, vol. 33, no. 3, pp. 305-315. https://doi.org/10.1097/CJI.0b013e3181c0c3cb
Vera, Juan F. ; Brenner, Lara J. ; Gerdemann, Ulrike ; Ngo, Minhtran C. ; Sili, Uluhan ; Liu, Hao ; Wilson, John ; Dotti, Gianpietro ; Heslop, Helen E. ; Leen, Ann M. ; Rooney, Cliona M. / Accelerated production of antigen-specific T cells for preclinical and clinical applications using gas-permeable rapid expansion cultureware (G-Rex). In: Journal of Immunotherapy. 2010 ; Vol. 33, No. 3. pp. 305-315.
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