Advantages of hydrophobic culture bags over flasks for the generation of monocyte-derived dendritic cells for clinical applications

Guyre, Cheryl A.; Fisher, Jan L.; Waugh, Mary G.; Wallace, Paul K.; Tretter, Christopher P.G.; Ernstoff, Marc S.; Barth, Richard J.

doi:10.1016/s0022-1759(02)00015-7

Cited by 28 publications

(28 citation statements)

References 24 publications

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“…Human DCs were generated using a standard protocol of leukapheresis, countercurrent elutriation, and differentiation of the monocyte fraction with GM-CSF and IL-4 for 7 d (Guyre et al 2002). The resulting immature DCs were split with half receiving LPS (2.5 mg/mL) and CD40L (10 mg/mL) for 24 h, and half left untreated.…”

Section: Resultsmentioning

confidence: 99%

“…Differentiation: Monocytes were incubated in VueLife bags in RPMI containing 10% FCS and GM-CSF (10 ng/mL) and IL-4 (20 ng/mL) for 7 d, receiving additional GM-CSF and IL-4 as above on day 4 (Guyre et al 2002). Maturation: iDCs were harvested, washed, and cultured in the presence of LPS (2.5 mg/mL) and CD40L (10 mg/mL), for 24 h at 37°C in VueLife bags.…”

Section: Generation and Maturation Of Dendritic Cellsmentioning

confidence: 99%

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Identification of TTP mRNA targets in human dendritic cells reveals TTP as a critical regulator of dendritic cell maturation

Emmons¹,

Townley-Tilson²,

Deleault³

et al. 2008

RNA

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Dendritic cells provide a critical link between innate and adaptive immunity and are essential to prime a naive T-cell response. The transition from immature dendritic cells to mature dendritic cells involves numerous changes in gene expression; however, the role of post-transcriptional changes in this process has been largely ignored. Tristetraprolin is an AU-rich element mRNAbinding protein that has been shown to regulate the stability of a number of cytokines and chemokines of mRNAs. Using TTP immunoprecipitations and Affymetrix GeneChips, we identified 393 messages as putative TTP mRNA targets in human dendritic cells. Gene ontology analysis revealed that ;25% of the identified mRNAs are associated with protein synthesis. We also identified six MHC Class I alleles, five MHC Class II alleles, seven chemokine and chemokine receptor genes, indoleamine 2,3 dioxygenase, and CD86 as putative TTP ligands. Real-time PCR was used to validate the GeneChip data for 15 putative target genes and functional studies performed for six target genes. These data establish that TTP regulates the expression of DUSP1, IDO, SOD2, CD86, and MHC Class I-B and F via the 39-untranslated region of each gene. A novel finding is the demonstration that TTP can interact with and regulate the expression of non-AU-rich element-containing messages. The data implicate TTP as having a broader role in regulating and limiting the immune response than previously suspected.

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Section: Resultsmentioning

confidence: 99%

Section: Generation and Maturation Of Dendritic Cellsmentioning

confidence: 99%

Identification of TTP mRNA targets in human dendritic cells reveals TTP as a critical regulator of dendritic cell maturation

Emmons¹,

Townley-Tilson²,

Deleault³

et al. 2008

RNA

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show abstract

“…Although for the clinical application of DCs the culture will need to take place in a fully enclosed system, for instance cell bags (Guyre et al 2002). Because of the simplicity of the protocol described here the transfer to such a system should be easily achieved.…”

Section: Discussionmentioning

confidence: 99%

Development of a standardized protocol for reproducible generation of matured monocyte-derived dendritic cells suitable for clinical application

Bohnenkamp

Noll

2003

Cytotechnology

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There is increasing interest in the generation of dendritic cells (DC) for cancer immunotherapy. In order to utilize DC in clinical trials it is necessary to have standardized, reproducible and easy to use protocols. We describe here the process development for the generation of DC as the result of investigation of culture conditions as well as consumption rates of medium and cytokines. Our studies demonstrate that highly viable DC (93 ± 2%) can be produced from CD14 + enriched monocytes via immunomagnetic beads in a high yield (31 ± 6%) with X-VIVO 15, 400 U ml À1 GM-CSF and 2000 U ml À1 IL-4 without serum and feeding. For the maturation of DC different cocktails (TNF-, IL-1, IL-6, PGE 2 and TNF-, PGE 2 ) were compared. In both cases cells expressed typical surface molecules of mature DC and induced high proliferative responses in mixed lymphocyte reactions which led to IFN-producing T-lymphocytes. The data suggest that the use of this optimized, easy to use protocol results in highly mature DC.

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“…In view of existing reports describing also the serumfree culture of DCs for clinical application [5,8,9] and because of potential disadvantages associated with the use of serum such as lot to lot variability and the presence of undefined serum factors, we considered DC culture in serumfree AIMV medium as an attractive alternative. However, from existing literature, we missed a conclusive direct comparison of such differently cultured DCs.…”

Section: Discussionmentioning

confidence: 99%

“…Both autologous as well as allogeneic serum derived from the patients or healthy donors, respectively, potentially introduce undefined ingredients with variable effects on DC function together with a lot to lot variability to the culture system. Serumfree culture of monocyte-derived DCs has been reported with media such as XVivo15, XVivo20 and AIMV [2][3][4][5][6][7][8][9][10]. From these reports, however, we could not deduce whether DCs cultured under such conditions are functionally equivalent to DCs cultured in the presence of human serum.…”

Section: Introductionmentioning

confidence: 87%

Functional characterization of monocyte-derived dendritic cells generated under serumfree culture conditions

et al. 2005

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Advantages of hydrophobic culture bags over flasks for the generation of monocyte-derived dendritic cells for clinical applications

Cited by 28 publications

References 24 publications

Identification of TTP mRNA targets in human dendritic cells reveals TTP as a critical regulator of dendritic cell maturation

Identification of TTP mRNA targets in human dendritic cells reveals TTP as a critical regulator of dendritic cell maturation

Development of a standardized protocol for reproducible generation of matured monocyte-derived dendritic cells suitable for clinical application

Functional characterization of monocyte-derived dendritic cells generated under serumfree culture conditions

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