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dendritic cell line, in vitro research
Abstract:
Research in vitro facilitates discovery, screening, and pilot experiments, often preceding
research in vivo. Several technical difficulties render Dendritic Cell (DC) research particularly
challenging, including the low frequency of DC in vivo, thorough isolation requirements,
and the vulnerability of DC ex vivo. Critically, there is not as yet a widely accepted human or
murine DC line and in vitro systems of DC research are limited. In this study, we report the
generation of new murine DC lines, named MutuDC, originating from cultures of splenic
CD8a conventional DC (cDC) tumors. By direct comparison to normalWT splenic cDC subsets,
we describe the phenotypic and functional features of the MutuDC lines and show
that they have retained all the major features of their natural counterpart in vivo, the splenic
CD8a cDC. These features include expression of surface markers Clec9A, DEC205, and
CD24, positive response to TLR3 and TLR9 but not TLR7 stimuli, secretion of cytokines,
and chemokines upon activation, as well as cross-presentation capacity. In addition to the
close resemblance to normal splenic CD8a cDC, a major advantage is the ease of derivation
and maintenance of the MutuDC lines, using standard culture medium and conditions,
importantly without adding supplementary growth factors or maturation-inducing stimuli
to the medium. Furthermore, genetically modified MutuDC lines have been successfully
obtained either by lentiviral transduction or by culture of DC tumors originating from genetically
modified mice. In view of the current lack of stable and functional DC lines, these
novel murine DC lines have the potential to serve as an important auxiliary tool for DC
research.
