The cryopreservation of high concentrated PBMC for dendritic cell (DC)-based cancer immunotherapy

Heo, Yoon Jeong; Son, Cheol; Chung, Joo-Seop; Park, You-Soo; Son, Jeong Hwa

doi:10.1016/j.cryobiol.2008.12.006

Cited by 17 publications

(11 citation statements)

References 32 publications

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“…As expected, the induction of maturation by poly(I/C) resulted in statistically significant differences in CD83 expression between immature and mature DC [43]. Therefore, our results confirm an earlier study that also reported no phenotypical differences between NC-DC and DC generated from CRF-PBMC [38]. …”

Section: Discussionsupporting

confidence: 91%

“…Our finding that total protein per plate of IPA-PBMC is already significantly reduced 24 h after culture of adherent cells led to the hypothesis that fewer IPA-PBMC recover compared with CRF-PBMC. It was shown that viability of cells after standard freezing procedures was significantly reduced [38]. Another contributing factor for reduced IPA-iDC numbers might be cryopreservation-dependent downregulation of adherence molecules on monocytes and therefore an increased loss of CD14+ cells during pre-plating.…”

Section: Discussionmentioning

confidence: 99%

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Controlled-rate freezer cryopreservation of highly concentrated peripheral blood mononuclear cells results in higher cell yields and superior autologous T-cell stimulation for dendritic cell-based immunotherapy

Buhl

Legler

Rosenberger

et al. 2012

Cancer Immunol Immunother

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Availability of large quantities of functionally effective dendritic cells (DC) represents one of the major challenges for immunotherapeutic trials against infectious or malignant diseases. Low numbers or insufficient T-cell activation of DC may result in premature termination of treatment and unsatisfying immune responses in clinical trials. Based on the notion that cryopreservation of monocytes is superior to cryopreservation of immature or mature DC in terms of resulting DC quantity and immuno-stimulatory capacity, we aimed to establish an optimized protocol for the cryopreservation of highly concentrated peripheral blood mononuclear cells (PBMC) for DC-based immunotherapy. Cryopreserved cell preparations were analyzed regarding quantitative recovery, viability, phenotype, and functional properties. In contrast to standard isopropyl alcohol (IPA) freezing, PBMC cryopreservation in an automated controlled-rate freezer (CRF) with subsequent thawing and differentiation resulted in significantly higher cell yields of immature and mature DC. Immature DC yields and total protein content after using CRF were comparable with results obtained with freshly prepared PBMC and exceeded results of standard IPA freezing by approximately 50 %. While differentiation markers, allogeneic T-cell stimulation, viability, and cytokine profiles were similar to DC from standard freezing procedures, DC generated from CRF-cryopreserved PBMC induced a significantly higher antigen-specific IFN-γ release from autologous effector T cells. In summary, automated controlled-rate freezing of highly concentrated PBMC represents an improved method for increasing DC yields and autologous T-cell stimulation.

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Section: Discussionsupporting

confidence: 91%

Section: Discussionmentioning

confidence: 99%

Controlled-rate freezer cryopreservation of highly concentrated peripheral blood mononuclear cells results in higher cell yields and superior autologous T-cell stimulation for dendritic cell-based immunotherapy

Buhl

Legler

Rosenberger

et al. 2012

Cancer Immunol Immunother

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“…The morphological features of DCs obtained from CD14 magnetic labeling were comparable to those of DCs obtained by Duddy and colleagues using the same protocols (Duddy et al, ) and to those of DCs obtained by Heo and colleagues using plastic adherence of monocytes (Heo et al, ).…”

Section: Discussionsupporting

confidence: 77%

Generation of tumor-specific cytotoxic T-lymphocytes from the peripheral blood of colorectal cancer patients for adoptive T-cell transfer

et al. 2015

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This study designs a strategy for an adoptive cellular therapy (ACT) protocol based on the ex-vivo selection of autologous peripheral blood-derived CD8-enriched T-cells, stimulated with dendritic cells (DCs) that had been pulsed with apoptotic tumor cells to generate cytotoxic T lymphocytes (CTLs) with anti-tumor activity. Seventy-eight colorectal cancer (CRC) patients were enrolled in this study. Tumor tissues and peripheral blood (PB) were obtained at surgery. Tissues were mechanically dissociated and cultured to obtain a primary tumor cell line from each patient. DCs were derived from peripheral blood mononuclear cells (PBMCs) using magnetic positive selection of CD14+ monocytes. Anti-tumor CTLs were elicited in co-/micro-cultures using DCs as antigen-presenting cells, autologous apoptotic tumor cells as a source of antigens, and CD8+ T lymphocytes as effectors. Interferon-γ (IFN-γ) secretion was assessed by ELISpot assays to evaluate the activation of the CTLs against the autologous tumor cells. Primary tumor cell lines were obtained from 20 of 78 patients (25.6%). DCs were generated from 26 patients, and of them, corresponding tumor cell lines were derived from six patients. ELISpot results showed that significant IFN-γ secretion was detected after different numbers of stimulations for two patients, whereas weak secretion was observed for three patients. Despite difficulties due to contamination of several primary tumor cell lines with gut intestinal flora, the results suggest that the generation of tumor-specific CTLs is feasible from patients with CRC, and could be useful for supporting an ACT approach in CRC.

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“…However, differences have been reported in the freezing of PBMCs with respect to viability and cell populations by many researchers [23,25,26]. On the basis of this literature, the Immunology of Diabetes Society T-Cell Workshop Committee decided to identify an optimal freezing protocol that would demonstrate minimal changes from responses to fresh cells but also a protocol that would allow for the highest and most reproducible responses to be obtained in assays currently being utilized in T1D research studies [6,10,27].…”

Section: Discussionmentioning

confidence: 98%

Comparison of cryopreservation methods on T‐cell responses to islet and control antigens from type 1 diabetic patients and controls

Brooks-Worrell¹,

Tree²,

Mannering³

et al. 2011

Diabetes Metabolism Res

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The cryopreservation of high concentrated PBMC for dendritic cell (DC)-based cancer immunotherapy

Cited by 17 publications

References 32 publications

Controlled-rate freezer cryopreservation of highly concentrated peripheral blood mononuclear cells results in higher cell yields and superior autologous T-cell stimulation for dendritic cell-based immunotherapy

Controlled-rate freezer cryopreservation of highly concentrated peripheral blood mononuclear cells results in higher cell yields and superior autologous T-cell stimulation for dendritic cell-based immunotherapy

Generation of tumor-specific cytotoxic T-lymphocytes from the peripheral blood of colorectal cancer patients for adoptive T-cell transfer

Comparison of cryopreservation methods on T‐cell responses to islet and control antigens from type 1 diabetic patients and controls

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