Maintaining dendritic cell viability in culture.

Abstract:

:When mouse dendritic cells (DCs) are isolated from tissues, purified and placed in a nutritive culture they die more rapidly than would be expected from their normal turnover in vivo. This can distort culture assays of DC function. We therefore tested several approaches to prolonging DC survival in culture. Of several cytokines tested granulocyte-macrophage colony stimulating factor was most effective at preserving the viability of conventional DCs (cDCs) but was ineffective for plasmacytoid DCs (pDCs). Surprisingly, Fms-like tyrosine kinase 3 ligand, crucial for DC development, produced only a marginal improvement in DC survival in culture, and interleukin-3, reported to prevent apoptosis of human pDCs, produced only a minor improvement in survival of mouse DCs. Genetic manipulation of cell death pathways was also tested, to avoid activation effects exerted by cytokine signalling. The isolation of DCs from mice overexpressing Bcl-2 was especially effective in maintaining pDC viability but gave a lesser improvement in cDC viability. DCs isolated from Bim(-/-)Noxa(-/-) mice also showed improved culture survival, but in this case with pDCs showing the least improvement.

journal_name

Mol Immunol

journal_title

Molecular immunology

authors

Vremec D,Hansen J,Strasser A,Acha-Orbea H,Zhan Y,O'Keeffe M,Shortman K

doi

10.1016/j.molimm.2014.07.011

subject

Has Abstract

pub_date

2015-02-01 00:00:00

pages

264-7

issue

2

eissn

0161-5890

issn

1872-9142

pii

S0161-5890(14)00181-3

journal_volume

63

pub_type

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