Penicillin and streptomycin

U2OS (human bone osteosarcoma cell line) and Hela (human cervical adenocarcinoma epithelial cell line) were gifts from Prof. R.-H. Chen’s laboratory (Academia Sinica, Taipei, Taiwan) and were maintained in DMEM-high glucose (Invitrogen) supplemented with 10% FBS (Invitrogen) and 1% antibiotic solution (penicillin and streptomycin; Invitrogen) under 5% CO₂. HFF1 (human foreskin fibroblasts) cells were purchased from ATCC and were maintained in DMEM-high glucose supplemented with 15% FBS and 1% antibiotic solution (penicillin and streptomycin) under 5% CO₂. Human mesenchymal stem cells (MSCs) were purchased from Lonza and were maintained in DMEM-low glucose supplemented with 10% FBS (Hyclone) and 1% antibiotic solution (penicillin and streptomycin) under 5% CO₂.
The homo plasma was obtained from human blood donated by blood donors. All methods related to human blood were carried out in accordance with relevant guidelines and regulations. All experiments protocols related to human blood were approved by the Ethics Committee of the Institutional Review Board (IRB) of National Yang-Ming University. Informed consent was obtained from all subjects. The porcine plasma was obtained from CHAISHAN FOODS CO., LTD.

GBM cell lines U87-MG, LN-18, and SF-188 were cultured in 1:1 DMEM/F12 (GIBCO, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (Corning, Manassas, VA, USA) and 1% penicillin and streptomycin. GBM cell line SJ-GBM2 was cultured in Iscove’s Modified Dulbecco’s Media (GIBCO, Carlsbad, CA, USA) supplemented with 20% fetal bovine serum, Insulin-Transferrin-Selenium supplement (Corning, Bedford, MA, USA), and 1% penicillin and streptomycin (ATCC, Manassas, VA, USA). Cell cultures were maintained at 37 °C in humidified air containing 5% CO₂.

PC12 Tet-Off cells (RRID:CVCL_V361) (Clontech, 631134, termed PC12 cells for simplicity) were maintained in DMEM medium (1 g/L glucose, Lonza) supplemented with 10% FBS, 5% horse serum (HS, Thermo Fisher Scientific Inc.), and 100 U/ml each of penicillin and streptomycin (Thermo Fisher Scientific Inc., Cat. No. 15140-122) and 2 mM GlutaMAX (Thermo Fisher Scientific Inc., Cat. No. 35050038). Starvation of PC12 cells was induced by 1% FCS, 100 U/ml each of penicillin and streptomycin and 2 mM GlutaMAX, but no HS. Cell-based assays for PC12 cells were performed in starvation conditions. A549 cells (RRID:CVCL_0023) (ATCC, CCL-185) were cultured in DMEM medium (4.5 g/L glucose) supplemented with 10% FCS and 100 U/ml each of penicillin and streptomycin and 2 mM GlutaMAX. T-47D cells (RRID:CVCL_0553) (ATCC, HTB-133) were cultured in RPMI 1640 medium supplemented with human insulin (f.c. 125 μg/L) (Sigma-Aldrich), 10% FCS and 100 U/ml each of penicillin and streptomycin and 2 mM GlutaMAX.

Primary mouse bone marrow-derived macrophages (BMDMs) were generated from the bone marrow of wild type and the indicated mutant mice. Cells were grown for 5–6 days in IMDM (Thermo Fisher Scientific, 12440053) supplemented with 1% non-essential amino acids (Thermo Fisher Scientific, 11140–050), 10% FBS (Biowest, S1620), 30% L929 conditioned media, and 1% penicillin and streptomycin (Thermo Fisher Scientific, 15070–063). BMDMs were then seeded into antibiotic-free media at a concentration of 1 × 10⁶ cells into 12-well plates and incubated overnight. The human monocytic cell line THP-1 (ATCC, TIB-202) was cultured in RPMI media (Corning, 10–040-CV) supplemented with 10% FBS and 1% penicillin and streptomycin. The primary umbilical vein endothelial cells from normal human (HUVEC) (ATCC, PCS-100–013) were cultured in vascular cell basal medium (ATCC, PCS-100–030) containing cell growth factors (ATCC, PCS-100–040) and 1% penicillin and streptomycin.

Primary CD14+ monocytes were isolated from fresh venous blood, obtained from healthy donors, using Lymphoprep (Stemcell Technologies) density gradient followed by magnetic cell sorting with CD14+‏ magnetic beads (Miltenyi Biotec). The cells were cultured in X-Vivo15 media (Lonza) supplemented with 2.25 mM L-glutamine at 37⁰C in 5% CO2 (Fortunato, 2014 (link)). Primary human foreskin fibroblasts (HFF) (ATCC CRL-1634) were maintained in DMEM with 10% fetal bovine serum (FBS), 2 mM L-glutamine, and 100 units/ml penicillin and streptomycin (Beit-Haemek, Israel).
The TB40E virus containing an SV40-GFP tag (TB40E-GFP) was described previously (O'Connor and Murphy, 2012 (link); Sinzger et al., 2008b (link)). Virus was propagated by electroporation of infectious bacterial artificial chromosome (BAC) DNA into fibroblasts using the Amaxa P2 4D-Nucleofector kit (Lonza) according to the manufacturer’s instructions. Viral stocks were concentrated by centrifugation at 26000xg, 4⁰C for 120 min. Infectious virus yields were assayed on THP-1 cells (ATCC TIB-202).