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Titolo:
An advanced culture method for generating large quantities of highly pure dendritic cells from mouse bone marrow
Autore:
Lutz, MB; Kukutsch, N; Ogilvie, ALJ; Rossner, S; Koch, F; Romani, N; Schuler, G;
Indirizzi:
Univ Erlangen Nurnberg, Dept Dermatol, D-91052 Erlangen, Germany Univ Erlangen Nurnberg Erlangen Germany D-91052 -91052 Erlangen, Germany Innsbruck Univ, Dept Dermatol, A-6020 Innsbruck, Austria Innsbruck Univ Innsbruck Austria A-6020 matol, A-6020 Innsbruck, Austria
Titolo Testata:
JOURNAL OF IMMUNOLOGICAL METHODS
fascicolo: 1, volume: 223, anno: 1999,
pagine: 77 - 92
SICI:
0022-1759(19990201)223:1<77:AACMFG>2.0.ZU;2-D
Fonte:
ISI
Lingua:
ENG
Soggetto:
COLONY-STIMULATING FACTOR; TNF-ALPHA; GM-CSF; CYTOKINES; ANTIGEN; BLOOD; DIFFERENTIATION; PROGENITORS; EXPRESSION; PRECURSORS;
Keywords:
dendritic cells; bone marrow; GM-CSF; culture method; mouse;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
26
Recensione:
Indirizzi per estratti:
Indirizzo: Lutz, MB Univrmanyngen Nurnberg, Dept Dermatol, Hartmannstr 14, D-91052 Erlangen, Ge Univ Erlangen Nurnberg Hartmannstr 14 Erlangen Germany D-91052 e
Citazione:
M.B. Lutz et al., "An advanced culture method for generating large quantities of highly pure dendritic cells from mouse bone marrow", J IMMUNOL M, 223(1), 1999, pp. 77-92

Abstract

As dendritic cells (DC) are rare populations in all organs, their generation from hematopoietic precursors in large quantities has proven critical tostudy their biology. From murine bone marrow about 5 X 10(6) cells at 70% purity are obtained per mouse after 8 days of culture with GM-CSF. We have improved this standard method and routinely achieve a 50-fold higher yield,i.e., 1-3 X 10(8) immature and mature DC per mouse at 90-95% purity. The major modifications were: (i) the avoidance of any active depletion of bone marrow cell subpopulations to circumvent loss of precursors, (ii) a lower plating density of bone marrow cells, (iii) a prolonged culture period of 10-12 days, (iv) the reduction of the GM-CSF dose from day 8 or 10 onwards toreduce granulocyte contaminations. The final non-adherent population at day 10-12 constitutes a mixture of immature and mature DC. Further maturationof DC could be induced by high doses of LPS or TNF-alpha for the last 24 h, where 50-70% of the non-adherent fraction represented mature DC with highlevels of NLDC-145, CD86 and CD40. This method allows by simple means the generation of high numbers of murine DC with very low B cell or granulocytecontaminations. It will be valuable to study DC biology notably at the molecular level. (C) 1999 Elsevier Science B.V. All rights reserved.

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Documento generato il 31/10/20 alle ore 00:09:32