Culture of human T cells in stirred bioreactors for cellular immunotherapy applications: Shear, proliferation, and the IL-2 receptor

Carswell, Kathleen S.; Papoutsakis, Eleftherios T.

doi:10.1002/(sici)1097-0290(20000505)68:3<328::aid-bit11>3.0.co;2-v

Cited by 57 publications

(43 citation statements)

References 40 publications

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“…A low agitation rate has little effect on the expression of surface molecules and receptors on CD34 + HSPCs It was documented that the hydrodynamic forces produced by agitation influenced the transcriptional levels of cell surface receptors and changed the physiological response and proliferative potential, implying hydrodynamic force is an important operational parameter in the art of cell culture [5,6]. Furthermore, the proliferation of hematopoietic cells is dependent on the agitation rate and impellor configuration [2].…”

Section: Cell Mol Biol Lett 482mentioning

confidence: 99%

A comparative gene-expression analysis of CD34+ hematopoietic stem and progenitor cells grown in static and stirred culture systems

Liu

Cai

et al. 2006

Cellular and Molecular Biology Letters

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Static and stirred culture systems are widely used to expand hematopoietic cells, but differential culture performances are observed between these systems. We hypothesize that these differential culture outcomes are caused by the physiological responses of CD34 + hematopoietic stem and progenitor cells (HSPCs) to the different physical microenvironments created in these culture devices. To understand the genetic changes provoked by culture microenvironments, the gene expression profiling of CD34 + HSPCs grown in static and stirred culture systems was compared using SMART-PCR and cDNA arrays. The results revealed that 103 and 99 genes were significantly expressed in CD34 + cells from static and stirred systems, respectively. Of those, 91 have similar levels of expression, while 12 show differential transcription levels. These differentially expressed genes are mainly involved in anti-oxidation, DNA repair, apoptosis, and chemotactic activity. A quantitative molecular understanding of the influences of growth microenvironments on transcriptional events in CD34 + HSPCs should give new insights into optimizing culture strategies to produce hematopoietic cells.

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Section: Cell Mol Biol Lett 482mentioning

confidence: 99%

A comparative gene-expression analysis of CD34+ hematopoietic stem and progenitor cells grown in static and stirred culture systems

Liu

Cai

et al. 2006

Cellular and Molecular Biology Letters

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“…To determine optimum conditions for expanding CMVspecific CTL in suspension culture, we conducted expansion experiments using PHA-stimulated lymphocytes in spinner flasks as described previously (Carswell and Papoutsakis, 2000). Briefly, PBMC were isolated and resuspended in TCM at 1 Â 10 6 cells/mL and stimulated with 5 Ag/mL PHA for 5 days.…”

Section: Expansion Of Pha-stimulated T Cells In Suspension Culturementioning

confidence: 99%

“…There has been interest in recent years in developing methods for large-scale expansion of Ag-specific CTL using a variety of expansion techniques and culture vessels, such as T-flasks, gas-permeable tissue culture bags, artificial capillary bioreactors, and suspension bioreactors (Bohnenkamp et al, 2002;Carswell and Papoutsakis, 2000;Freedman et al, 1994;Levine et al, 1998;Pierson et al, 1996;Robinet et al, 1998). Investigation into the potential of stirred suspension bioreactor systems shows promise for a number of reasons.…”

Section: Introductionmentioning

confidence: 99%

“…Finally, due to the large cell numbers required for adoptive immunotherapy trials, suspension cultures are readily scaled up to meet higher culture volumes (an estimated 20-L working volume is required to achieve cell numbers of 10 10 at modest cell densities of 5 Â 10 6 cells/mL). Preliminary investigations into the use of stirred suspension reactors for ex vivo expansion of lymphocytes indicate that this reactor type might be applied to Ag-specific CTL expansion (Carswell and Papoutsakis, 2000).…”

Section: Introductionmentioning

confidence: 99%

“…Although these initial studies were successful in expanding lymphocytes, the data reflect the expansion of peripheral blood mononuclear cells (Bohnenkamp et al, 2002;Carswell and Papoutsakis, 2000), T cells derived from malignant biopsies (Freedman et al, 1994), or patients with viral infection (Levine et al, 1998) rather than in vitro-selected Ag-specific CTL. There has been much interest in selecting Ag-specific CTL using potent antigen-presenting dendritic cells (DC) pulsed with MHC class I peptide epitopes (nonamer peptides) to reduce the frequency of allogeneic-reactive T cells within the culture population.…”

Section: Introductionmentioning

confidence: 99%

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Large‐scale expansion of cytomegalovirus‐specific cytotoxic T cells in suspension culture

Foster¹,

Forrester²,

Gottlieb³

et al. 2003

Biotech & Bioengineering

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Clinical trials in recent years involving the adoptive transfer of antigen-specific cytotoxic T lymphocytes (CTL) have shown promise in restoring immunity against viral infection and reducing tumor burden in patients with solid and hematological malignancies. However, the large cell number required to achieve efficacy, 10(9) to 10(11), makes routine application of adoptive immunotherapy impractical. Investigation into new methods of CTL expansion may be useful in addressing this problem. Use of stirred suspension bioreactors are one such method that may allow large-scale T-cell expansion. Suspension cultures offer advantages over conventional static culture methods, including providing a homogeneous culture environment, and the potential for optimization and control of culture conditions. We generated cytomegalovirus (CMV)-specific CTL and investigated the potential of stirred bioreactor systems for expansion of large cell numbers. We found that CTL can be readily expanded ( > 200-fold) from cryopreserved stocks by nonspecific stimulation in the presence of allogeneic feeder cells and interleukin-2 (IL-2). Activated CTL inoculated into either suspension or static cultures could be subsequently expanded tenfold, and showed similar growth kinetics and metabolism independent of the culture vessel used. Furthermore, CTL remained specific for CMVpp65 peptide through the expansion phases, as demonstrated by pp65-tetramer staining ( > 95% tetramer(+)) and cytotoxicity assays. This study indicates that suspension reactor systems may be useful in large-scale expansion of antigen-specific CTL lines or clones, and may facilitate the advancement of routine adoptive immunotherapy.

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Bioprocess Development for the Cultivation of Human T-lymphocytes

et al. 2001

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Culture of human T cells in stirred bioreactors for cellular immunotherapy applications: Shear, proliferation, and the IL-2 receptor

Cited by 57 publications

References 40 publications

A comparative gene-expression analysis of CD34+ hematopoietic stem and progenitor cells grown in static and stirred culture systems

A comparative gene-expression analysis of CD34+ hematopoietic stem and progenitor cells grown in static and stirred culture systems

Large‐scale expansion of cytomegalovirus‐specific cytotoxic T cells in suspension culture

Bioprocess Development for the Cultivation of Human T-lymphocytes

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