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Titolo:
Development of a clinical-scale method for generation of dendritic cells from PBMC for use in cancer immunotherapy
Autore:
Wong, ECC; Maher, VE; Hines, K; Lee, J; Carter, CS; Goletz, T; Kopp, W; Mackall, CL; Berzofsky, JA; Read, EJ;
Indirizzi:
NCI, Warren Grant Magnuson Clin Ctr, Dept Transfus Med, NIH, Bethesda, MD 20892 USA NCI Bethesda MD USA 20892 Dept Transfus Med, NIH, Bethesda, MD 20892 USA NCI, Metab Branch, NIH, Bethesda, MD USA NCI Bethesda MD USANCI, Metab Branch, NIH, Bethesda, MD USA NCI, Pediat Oncol Branch, NIH, Bethesda, MD USA NCI Bethesda MD USANCI, Pediat Oncol Branch, NIH, Bethesda, MD USA SAIC, Frederick, MD USA SAIC Frederick MD USASAIC, Frederick, MD USA
Titolo Testata:
CYTOTHERAPY
fascicolo: 1, volume: 3, anno: 2001,
pagine: 19 - 29
SICI:
1465-3249(2001)3:1<19:DOACMF>2.0.ZU;2-R
Fonte:
ISI
Lingua:
ENG
Soggetto:
BLOOD MONOCYTES; SERUM; ANTIGEN; MATURE;
Keywords:
dendritic cells (DCs); peripheral blood monocytes; immunotherapy;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Clinical Medicine
Life Sciences
Citazioni:
19
Recensione:
Indirizzi per estratti:
Indirizzo: Read, EJ NCI, Warren Grant Magnuson Clin Ctr, Dept Transfus Med, NIH, Bethesda, MD 20892 USA NCI Bethesda MD USA 20892 nsfus Med, NIH, Bethesda, MD 20892 USA
Citazione:
E.C.C. Wong et al., "Development of a clinical-scale method for generation of dendritic cells from PBMC for use in cancer immunotherapy", CYTOTHERAPY, 3(1), 2001, pp. 19-29

Abstract

Background There is growing interest bt the use of dendritic cells (DCs) for treatment of malignancy and infectious disease Our goal was to develop aclinical-scale method to prepare autologous DCs for cancer clinical trialsMethods PBMC were collected from normal donors or cancer patients by automated leukapheresis, purified by counterflow centrifugal elutriation and placed into culture in polystyrene flasks at 1 X 10(6) cells/mL for 5-7 days at 37 degreesC, with 5% CO2, with IL-4 and GM-CSF. Conditions investigated included media formulation, supplementation with heat-inactivated allogeneicAB serum or autologous plasma and time to harvest (Day 5 or Day 7). DCs were evaluated for morphology, quantitative yield viability phenotype and function, including mixed leukocyte response and recall response to tetanus toroid and influenza virus. Results DCs with a typical immature phenotype (CD14-negative, CD1a-positive, mannose receptor-positive, CD80-positive, CD83-negntive) were generated most consistently in RPMI 1640 supplemented with 10% allogeneic AB serum or10% autologous plasma. Cell yield was higher at Day 5 than Day 7, without detectable differences in phenotype or function. In pediatric sarcoma patients autologous DCs bad enhanced function compared with monocytes from whichthey were generated. In this patient group, starting with 8.0 +/- 3.7 x 10(8 fr)esh or cryopreserved autologous monocytes, DC yield was 2.1 +/- 1.0 x10(8) cells, or 29% of the starting monocyte number:Discussion In the optimized clinical-scale method, purified peripheral monocytes are cultured for 5 days in flasks at 1 X 10(6) cells/mL in RPMI 1640, 10% allogeneic AB serum or autologous plasma, IL-4 and CM-CSF: This method avoids the use of FBS and results in in immature DCs suitable for clinical trials.

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Documento generato il 05/04/20 alle ore 06:50:18