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Direct Reprogramming of Mouse Embryonic Fibroblasts to Conventional Type 1 Dendritic Cells by Enforced Expression of Transcription Factors

Rosa, Fábio F LU ; Pires, Cristiana F LU ; Zimmermannova, Olga LU and Pereira, Carlos-Filipe LU orcid (2020) In Bio-protocol 10(10).
Abstract

Ectopic expression of transcription factor combinations has been recently demonstrated to reprogram differentiated somatic cells towards the dendritic cell (DC) lineage without reversion to a multipotent state. DCs have the ability to induce potent and long-lasting adaptive immune responses. In particular, conventional type 1 DCs (cDC1s) excel on antigen cross-presentation, a critical step for inducing CD8+ T cell cytotoxic responses. The rarity of naturally occurring cDC1s and lack of in vitro methodologies for the generation of pure cDC1 populations strongly hinders the study of cDC1 lineage specification and function. Here, we describe a protocol for the generation of induced DCs (iDCs) by lentiviral-mediated expression of the... (More)

Ectopic expression of transcription factor combinations has been recently demonstrated to reprogram differentiated somatic cells towards the dendritic cell (DC) lineage without reversion to a multipotent state. DCs have the ability to induce potent and long-lasting adaptive immune responses. In particular, conventional type 1 DCs (cDC1s) excel on antigen cross-presentation, a critical step for inducing CD8+ T cell cytotoxic responses. The rarity of naturally occurring cDC1s and lack of in vitro methodologies for the generation of pure cDC1 populations strongly hinders the study of cDC1 lineage specification and function. Here, we describe a protocol for the generation of induced DCs (iDCs) by lentiviral-mediated expression of the transcription factors PU.1, IRF8 and BATF3 in mouse embryonic fibroblasts. iDCs acquire DC morphology, cDC1 phenotype and transcriptional signatures within 9 days. iDCs generated with this protocol acquire functional ability to respond to inflammatory stimuli, engulf dead cells, process and cross-present antigens to CD8+ T cells. DC reprogramming provides a simple and tractable system to generate high numbers of cDC1-like cells for high content screening, opening new avenues to better understand cDC1 specification and function. In the future, faithful induction of cDC1 fate in fibroblasts may lead to the generation of patient-specific DCs for vaccination.

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author
; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Bio-protocol
volume
10
issue
10
article number
e3619
publisher
Bio-protocol LLC
external identifiers
  • scopus:85103339983
  • pmid:33659292
ISSN
2331-8325
DOI
10.21769/BioProtoc.3619
language
English
LU publication?
yes
additional info
Copyright © 2020 The Authors; exclusive licensee Bio-protocol LLC.
id
3d3cca6c-6e10-40c6-8b7d-8a0c97957fff
date added to LUP
2022-12-08 15:00:33
date last changed
2024-04-05 17:40:15
@article{3d3cca6c-6e10-40c6-8b7d-8a0c97957fff,
  abstract     = {{<p>Ectopic expression of transcription factor combinations has been recently demonstrated to reprogram differentiated somatic cells towards the dendritic cell (DC) lineage without reversion to a multipotent state. DCs have the ability to induce potent and long-lasting adaptive immune responses. In particular, conventional type 1 DCs (cDC1s) excel on antigen cross-presentation, a critical step for inducing CD8+ T cell cytotoxic responses. The rarity of naturally occurring cDC1s and lack of in vitro methodologies for the generation of pure cDC1 populations strongly hinders the study of cDC1 lineage specification and function. Here, we describe a protocol for the generation of induced DCs (iDCs) by lentiviral-mediated expression of the transcription factors PU.1, IRF8 and BATF3 in mouse embryonic fibroblasts. iDCs acquire DC morphology, cDC1 phenotype and transcriptional signatures within 9 days. iDCs generated with this protocol acquire functional ability to respond to inflammatory stimuli, engulf dead cells, process and cross-present antigens to CD8+ T cells. DC reprogramming provides a simple and tractable system to generate high numbers of cDC1-like cells for high content screening, opening new avenues to better understand cDC1 specification and function. In the future, faithful induction of cDC1 fate in fibroblasts may lead to the generation of patient-specific DCs for vaccination.</p>}},
  author       = {{Rosa, Fábio F and Pires, Cristiana F and Zimmermannova, Olga and Pereira, Carlos-Filipe}},
  issn         = {{2331-8325}},
  language     = {{eng}},
  month        = {{05}},
  number       = {{10}},
  publisher    = {{Bio-protocol LLC}},
  series       = {{Bio-protocol}},
  title        = {{Direct Reprogramming of Mouse Embryonic Fibroblasts to Conventional Type 1 Dendritic Cells by Enforced Expression of Transcription Factors}},
  url          = {{http://dx.doi.org/10.21769/BioProtoc.3619}},
  doi          = {{10.21769/BioProtoc.3619}},
  volume       = {{10}},
  year         = {{2020}},
}