Ascorbic acid supports ex vivo generation of plasmacytoid dendritic cells from circulating hematopoietic stem cells

eLife

Albert Gris-Oliver¹  Anders Laustsen¹  Martin R Jakobsen¹  Sabina Sánchez Hernández¹  Rasmus O Bak²  Renée M van der Sluis²  Ena Cemalovic³  Lars Henning Pedersen⁴  Niels Uldbjerg⁵  Hai Q Tang⁶ 

[1] Department of Biomedicine, Aarhus University, Aarhus, Denmark;Department of Biomedicine, Aarhus University, Aarhus, Denmark;Aarhus Institute of Advanced Studies, Aarhus University, Aarhus, Denmark;Department of Biomedicine, Aarhus University, Aarhus, Denmark;Centre of Molecular Inflammation Research, Department of Clinical and Molecular Medicine, Norwegian University of Science and Technology, Trondheim, Norway;Clinic of Medicine, St. Olav's University Hospital, Trondheim, Norway;Department of Biomedicine, Aarhus University, Aarhus, Denmark;Department of Obstetrics and Gynaecology, Aarhus University Hospital, Aarhus, Denmark;Department of Clinical Medicine, Aarhus University, Aarhus, Denmark;Department of Clinical Medicine, Aarhus University, Aarhus, Denmark;Department of Obstetrics and Gynaecology, Aarhus University Hospital, Aarhus, Denmark;

关键词: ascorbic acid;    plasmacytoid;    pDC;    dendritic cells;    hematopoietic stem cells;    vitamin c;    ex;    vivo;    differentiation;    immunotherapy;    TLR7;    TLR9;    cHSPC;    Human;   

DOI  :  10.7554/eLife.65528

来源: eLife Sciences Publications, Ltd

【 摘 要 】

Plasmacytoid dendritic cells (pDCs) constitute a rare type of immune cell with multifaceted functions, but their potential use as a cell-based immunotherapy is challenged by the scarce cell numbers that can be extracted from blood. Here, we systematically investigate culture parameters for generating pDCs from hematopoietic stem and progenitor cells (HSPCs). Using optimized conditions combined with implementation of HSPC pre-expansion, we generate an average of 465 million HSPC-derived pDCs (HSPC-pDCs) starting from 100,000 cord blood-derived HSPCs. Furthermore, we demonstrate that such protocol allows HSPC-pDC generation from whole-blood HSPCs, and these cells display a pDC phenotype and function. Using GMP-compliant medium, we observe a remarkable loss of TLR7/9 responses, which is rescued by ascorbic acid supplementation. Ascorbic acid induces transcriptional signatures associated with pDC-specific innate immune pathways, suggesting an undescribed role of ascorbic acid for pDC functionality. This constitutes the first protocol for generating pDCs from whole blood and lays the foundation for investigating HSPC-pDCs for cell-based immunotherapy.

【 授权许可】

CC BY