The In Vitro Differentiation of Human CD141+CLEC9A+ Dendritic Cells from Mobilized Peripheral Blood CD34+ Hematopoietic Stem Cells.
Curr Protoc
; 2(4): e410, 2022 Apr.
Article
em En
| MEDLINE
| ID: mdl-35435334
ABSTRACT
As shown in various preclinical studies, conventional type-1 dendritic cells, or cDC1s, play a critical role in the immunological rejection of tumors and in the defense against pathogens. This indispensability stems from their potent capacity to activate cytotoxic T cells, especially via the cross-presentation of exogenous antigens. For this reason, cDC1s have become an attractive target for immunotherapy. Here we report a simplified method for generating large numbers of cDC1-like cells in vitro from mobilized human peripheral blood CD34+ hematopoietic stem cells using FMS-like tyrosine kinase 3 ligand (FLT3L) and granulocyte-macrophage colony-stimulating factor (GM-CSF). An important aspect of this Protocol is the growth of cells on a non-tissue culture-treated surface rather than on a tissue culture-treated surface since the latter suppresses cDC1-marker expression. The resulting CD11c+ DCs express high levels of cDC1-specific markers such as CD141, CLEC9A, TLR3, and several DC maturation markers. Compared to alternative differentiation methods, this method generates large numbers of cDC1-like cells without the need for immortalized feeder cells and should prove useful for studying cDC1 immunobiology and clinical applications of this DC subset. © 2022 Wiley Periodicals LLC. Basic Protocol Generation of human CD141+CLEC9A+ dendritic cells from mobilized peripheral blood CD34+ hematopoietic stem cells Support Protocol Flow cytometric immunophenotyping of CD141+ dendritic cells.
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Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Células Dendríticas
/
Apresentação Cruzada
Tipo de estudo:
Guideline
Limite:
Humans
Idioma:
En
Revista:
Curr Protoc
Ano de publicação:
2022
Tipo de documento:
Article