citations

                    Human CD34+-derived plasmacytoid dendritic cells as surrogates for primary pDCs and potential cancer immunotherapy
                

Authors:

Fiore G, Weckwarth W, Paetzold K, Albertí Servera L, Gies M, Rosenhauer J, Antoniolli M, Nassiri S, Schmeing S, Dettling S, Soni B, Majety M, Krug AB, Hoves S, Wolf MJ

In:

Source: Frontiers in Immunology
Publication Date: (2024)
Issue: 15: 1433119

Research Area:

Cancer Research/Cell Biology

Immunotherapy / Hematology

Basic Research

Molecular Biology

Regenerative medicine

Cells used in publication:

CD34+ cell, human: Species: human; Tissue Origin: blood

Experiment

2.1.2.2 Human CD34+ HSCs, peripheral blood mononuclear cells, and patient tumor samples

Cryopreserved human cord blood CD34+ hematopoietic stem and progenitor cells (HSCs) were purchased from Lonza.

2.1.2.3 Media

CD34+ HSCs were expanded in expansion medium: SFEM II stem pan medium (Stemcell Technologies) supplemented with 10% heatinactivated FCS, 1% sodium-pyruvate, 1% penicillin/streptomycin (all Anprotech), 100 ng/mL FLT3-L, 100 ng/mL SCF, 50 ng/mL TPO, and 5 ng/mL IL-7 (all Peprotech). Expanded CD34+ HSCs were further differentiated in differentiation medium: MS-5 medium supplemented with 100 ng/mL FLT3-L, 100 ng/mL SCF, 5 ng/mL IL-7 (Peprotech), 0.5mg/mL ascorbic acid (AA;MerckMillipore), 0.1 ng/mL GM-CSF (Biolegend), and 2.3 mM StemRegenin 1 (SR-1; Biogems).

Abstract

Introduction: Plasmacytoid dendritic cells (pDCs) are capable of triggering broad immune responses, yet, their scarcity in blood coupled to their reduced functionality in cancer, makes their therapeutic use for in situ activation or vaccination challenging.

Methods: We designed an in vitro differentiation protocol tailored for human pDCs from cord blood (CB) hematopoietic stem cells (HSCs) with StemRegenin 1 (SR-1) and GM-CSF supplementation. Next, we evaluated the identity and function of CB-pDCs compared to human primary pDCs. Furthermore, we tested the potential of CB-pDCs to support anti-tumor immune responses in co-culture with tumor explants from CRC patients.

Results: Here, we report an in vitro differentiation protocol enabling the generation of 200 pDCs per HSC and highlight the role of GM-CSF and SR-1 in CB-pDC differentiation and function. CB-pDCs exhibited a robust resemblance to primary pDCs phenotypically and functionally. Transcriptomic analysis confirmed strong homology at both, baseline and upon TLR9 or TLR7 stimulation. Further, we could confirm the potential of CB-pDCs to promote inflammation in the tumor microenvironment by eliciting cytokines associated with NK and T cell recruitment and function upon TLR7 stimulation ex vivo in patient tumor explants.

Discussion: This study highlights CB-pDCs as surrogates for primary pDCs to investigate their biology and for their potential use as cell therapy in cancer.

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