Penicillin streptomycin antibiotics mixture

Human iPSC line CREM003i-BU3C2 (11 ) originating from a blood sample of a 40-year-old male was kept at 36°C and 0.5% CO₂. The cells were cultured in 6-well plates coated with Matrigel (Corning), with StemFlex medium (Gibco) supplemented with Primocin (InvovGen). The media was changed 48 h after a passage and then every 24 h. Cells were passaged using STEMPRO EZPassage tool (Thermo Fisher Scientific) when 70–80% confluent. HEK293T and HeLa transformed cell lines were derived from human embryonic kidney and cervical cancer cells, respectively. Both were cultured at 36°C and 0.5% CO₂, in 10 cm plates, with DMEM medium (Gibco) supplemented with 10% FBS and penicillin–streptomycin antibiotics mixture (Gibco). Media was changed every 3–4 days.

Cell culture medium (RPMI 1640, Lot# 1720776), phosphate buffered saline (PBS), fetal bovine serum (FBS, Gibco^®, Lot# 07F2235K), Penicillin/Streptomycin antibiotics mixture (Gibco^®), and L-glutamine (Gibco^®) were purchased from Life Technologies (Stockholm, Sweden). Alamar blue (Lot #GF218932) was purchased from Thermo scientific. The LDH (lactate dehydrogenase) activity kit (MAK066, Lot #B2C300726V) and Phorbol 12-myristate 13-acetate (PMA) were purchased from Sigma-Aldrich (Stockholm, Sweden, and the Luminex assay (Bio-Plex^® Multiplex System, Lot #64020782) from Bio-Rad (Solna, Sweden). HNO₃ (67–69%, trace-metal grade, Lot #1113070) was bought from Fisher Chemical (Fischer Scientific, Loughborough, UK Limited).
A549 cells (human type II alveolar epithelial cell line) were obtained from NANoREG partners (Bundesanstalt Fuer Arbeitsschutz und Arbeitsmedizin, BAuA, Berlin, Germany) and THP-1 cells (peripheral blood monocyte cell line) were obtained from the American Type Culture Collection (Gaithersburg, USA).