Cell Lines and Peripheral Blood Mononuclear Cells

Adherent neuroblastoma cell lines derived from bone marrow metastases SK-N-DZ (DZ, passage 71 to 82, RRID:CVCL_1701), SK-N-SH (SH, passage 26 to 37, RRID:CVCL_0531), and SK-N-AS (AS, passage 2 to 7, RRID:CVCL_1700) were obtained from the ATCC and IGR-N-N91 (N91, passage 32 to 42, RRID:CVCL_8883) was from Institut Gustave Roussy Paris (Villejuif, France). Their identity was verified by HLA-A and B typing (PCR sequence-specific oligonucleotide method, EFS-AURA Grenoble, HLA laboratory). The cells were tested negative for Mycoplasma (MycoAlert Mycoplasma detection kit, Lonza). The neuroblastoma cell lines were cultured in DMEM high glucose supplemented with 10% FCS, 1× MEM Non-Essential amino acids, 20 μg/mL gentamicin (Thermo Fisher Scientific), referred to as complete medium at 37°C and 5% CO₂. Both DZ and N91 are MYCN-amplified cell lines, and AS and SH are MYCN-nonamplified.

Blood samples were collected from adult healthy volunteers (French national blood service, EFS) who gave written informed consent in accordance with the Declaration of Helsinki [cell collection approved by the French Ministry of Higher Education, Research and Innovation (Codecoh: GRE-DC-2019-3803)]. Peripheral blood mononuclear cells (PBMC) were isolated by density gradient on lymphocyte separation medium, and cryopreserved until use, by standard freezing methods [37.5% FCS, 10% DMSO (Sigma-Aldrich)].