title: 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.
authors: The Walter and Eliza Hall Institute, 1G Royal Parade, Parkville, Vic. 3052, Australia.
authors: David
authors: D
authors: Vremec
authors: The Walter and Eliza Hall Institute, 1G Royal Parade, Parkville, Vic. 3052, Australia.
authors: Jacinta
authors: J
authors: Hansen
authors: The Walter and Eliza Hall Institute, 1G Royal Parade, Parkville, Vic. 3052, Australia; Department of Medical Biology, University of Melbourne, Melbourne, Vic. 3010, Australia.
authors: Andreas
authors: A
authors: Strasser
authors: Department of Biochemistry, University of Lausanne, Epalinges, Switzerland.
authors: Hans
authors: H
authors: Acha-Orbea
authors: The Walter and Eliza Hall Institute, 1G Royal Parade, Parkville, Vic. 3052, Australia.
authors: Yifan
authors: Y
authors: Zhan
authors: Burnet Institute, Melbourne, Vic. 3004, Australia.
authors: Meredith
authors: M
authors: O'Keeffe
authors: The Walter and Eliza Hall Institute, 1G Royal Parade, Parkville, Vic. 3052, Australia; Department of Medical Biology, University of Melbourne, Melbourne, Vic. 3010, Australia; Burnet Institute, Melbourne, Vic. 3004, Australia. Electronic address: shortman@wehi.edu.au.
authors: Ken
authors: K
authors: Shortman
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D018929:Cell Culture Techniques / Q000379:methods*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D016207:Cytokines / Q000494:pharmacology; D003713:Dendritic Cells / Q000166:cytology* / Q000187:drug effects; D006801:Humans; D008810:Mice, Inbred C57BL; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics
chemical_list: D016207:Cytokines
keywords: Cytokines and DC survival; DC death; DC death pathways; pDC death