Comparison of monocyte enrichment by immuno-magnetic depletion or adherence for the clinical-scale generation of DC

Suen, Y; Lee, S.-M.; Aono, Frederick M.; Hou, Sheng; Loudovaris, Maureen; Ofstein, G.; Bender, James

doi:10.1080/146532401753277184

Cited by 37 publications

(27 citation statements)

References 24 publications

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“…Gradient centrifugation and (immuno)-magnetic activated cell sorting (MACS) are most commonly used [4]. Another technique, continuous counter-flow elutriation, separates cells into multiple fractions [5][6][7][8].…”

Section: Introductionmentioning

confidence: 99%

Isolation of monocytes from whole blood-derived buffy coats by continuous counter-flow elutriation

2006

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Monocytes (MOs) are the most commonly used precursors for the generation of dendritic cells (DCs) in vitro. Continuous counter-flow elutriation represents a promising tool to isolate MOs from white blood cell (WBC) products. Thirty whole blood-derived, AB0-identical buffy coats (BCs) were pooled using sterile technique (n = 5 experiments). For red blood cell (RBC) and polymorphonuclear cell (PMN) depletion, the BC pools were processed in a Cobe Spectra device (Gambro BCT) using the bone marrow program. Subsequently, continuous counter-flow elutriation in an Elutra device (Gambro BCT) was performed to enrich and purify MOs. BC pool volume averaged 1,260 +/- 14 ml containing 7.7 +/- 1.1 x 10(9) MOs. During 107 +/- 7 min, Cobe Spectra operation, the BC pools were processed for several times, and approximately 9,749 +/- 605 ml volume passed the device. Product volume and MO yield averaged 160 +/- 16 ml, and 4.3 +/- 1.3 x 10(9) cells, respectively. Elutra operation was performed within 59 +/- 0 min and yielded 2.5 +/- 0.9 x 10(9) MOs with a purity of 60 +/- 12%. Compared with the Cobe Spectra product cell count, MO recovery by Elutra averaged 59 +/- 10%. Elutriation of MOs from pooled BCs using Elutra exhibited comparatively low recovery and purity rates. This shortcoming may be due to the nature of the source material. Optimization of the elutriation procedure is necessary to improve MO enrichment from BCs.

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Section: Introductionmentioning

confidence: 99%

Isolation of monocytes from whole blood-derived buffy coats by continuous counter-flow elutriation

2006

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“…Monocytes are routinely obtained either by plastic adherence or by immunomagnetic selection of CD14 + cells from PB mononuclear cells. [13][14][15]17,18 However, the resulting monocyte preparations have been shown to contain variable numbers of contaminating lymphocytes. [17][18][19] Recently, it has been documented that DCs undergo rapid apoptosis during antigen-specific interaction with T cells both in vitro 20 and in vivo.…”

Section: Introductionmentioning

confidence: 99%

“…[13][14][15]17,18 However, the resulting monocyte preparations have been shown to contain variable numbers of contaminating lymphocytes. [17][18][19] Recently, it has been documented that DCs undergo rapid apoptosis during antigen-specific interaction with T cells both in vitro 20 and in vivo. [21][22][23][24] Several studies have shown that genetically modified DCs expressing TAA could effectively process and present antigenic peptides to T cells and trigger cytotoxic response against tumor cells expressing the target antigen.…”

Section: Introductionmentioning

confidence: 99%

Ex vivo generation of genetically modified dendritic cells for immunotherapy: implications of lymphocyte contamination

et al. 2005

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Genetically modified dendritic cell (DC) vaccines expressing tumor-associated antigens are currently used for cancer immunotherapy. Peripheral blood (PB) monocyte precursors are a relatively convenient source of DCs for use in clinical studies, but are often contaminated by lymphocytes. The current study was conducted to examine the impact of Tlymphocyte contamination on genetically modified DC product. PB monocyte-derived DCs were efficiently transduced (75-95%) with an HIV-1-based self-inactivating lentiviral vector encoding a model antigen, the enhanced green fluorescent protein (eGFP). The lymphocyte-free DC culture transduced with Lenti-eGFP showed stable expression of eGFP without measurable decline in viability. In contrast, the eGFP-positive DCs disappeared rapidly in transduced DC cultures containing lymphocyte contaminants, concurrent with detectable activation and expansion of T-lymphocytes. Upon antigen recall, these T cells elicited major histocompatability complex-restricted antigen-specific cytotoxicity against eGFP-positive autologous DCs and mitogen-stimulated T lymphoblasts, mainly through the perforin-mediated pathway. In summary, this study demonstrate that the relative purity of DC cultures could determine the persistence of gene-modified DC, which may affect the induction of effective immune responses by DC vaccination strategies. Gene Therapy (2005) 12, 259-271.

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“…In the first step GM-CSF and IL-4 promote the differentiation of immature monocyte derived DCs (iDCs) and in the second step pro-inflammatory factors induce the terminal maturation of DCs. [12][13]21,24,[26][27][28] The studies described here have two proposes, first to establish a low cost and highly efficient technique to obtain an enriched-CD14…”

Section: Discussionmentioning

confidence: 99%

“…[11][12][13] At present, most of the clinical trials use DCs generated from monocytes based on the findings of Sallusto and Lanzavechhia, 14 which consist in the generation of mature DCs by culturing peripheral blood mononuclear cells in the presence of GM-CSF plus interleukin 4 (IL-4) for one week followed by stimulation with TNF-α for 48h. GM-CSF is a growth factor for stem cells from granulocyte and macrophage lineages 15 and in DC cultures it appears to increase the expression of class II major histocompatibility complex (MHC) antigens, enhancing the antigen presenting capacity of DCs.…”

Section: Introductionmentioning

confidence: 99%

Maturation of dendritic cells following exposure to different maturational stimuli

Fontes¹,

Orellana²,

Palma³

et al. 2006

Rev. Bras. Hematol. Hemoter.

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Comparison of monocyte enrichment by immuno-magnetic depletion or adherence for the clinical-scale generation of DC

Cited by 37 publications

References 24 publications

Isolation of monocytes from whole blood-derived buffy coats by continuous counter-flow elutriation

Isolation of monocytes from whole blood-derived buffy coats by continuous counter-flow elutriation

Ex vivo generation of genetically modified dendritic cells for immunotherapy: implications of lymphocyte contamination

Maturation of dendritic cells following exposure to different maturational stimuli

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