U343mg

Normal Human Astrocytes (NHA) are normal human cells derived from healthy brain tissue, which were grown in astrocyte basal medium (ABM^TM) supplemented with astrocyte growth medium AGM^TM SingleQuots KIT (Lonza). U87MG, U251MG, and U343MG, glioblastoma cancer cell lines, were obtained from CLS Cell Lines Service GmbH (Eppelheim, Germany) cultured in DMEM (Dulbecco's Modified Eagle's Medium) supplemented with 10% heat inactivated fetal bovine serum, 1% L-Glutamine, 1% Sodium Pyruvate, 1% non-essential amino acid (Lonza), and 0.1% plasmocin TM prophylactic (InvivoGen). GBM 18 and GBM 63, primary cell lines of glioblastoma, were cultured in recommended medium DMEM/F-12 Ham (Sigma) supplemented with 15% heat inactivated fetal bovine serum, 2% L-Glutamine, 1% Sodium Pyruvate 1% non-essential (Lonza), 30% D-Glucose, and 1% antibiotic mixture, at 37°C in a humidified atmosphere with 5% carbon dioxide. The adherent primary cultures of brain tumor cells (designated as GBMn) were isolated accordingly to the procedure previously described by our group (13 (link)).

Expression of ZIKV E proteins was carried out using a modified version of pcDNA3.1 (pME) as described earlier [8 (link),27 (link),30 (link)] and the pSVATGrev plasmid [31 (link)]. Green fluorescent protein (GFP) was expressed by plasmid pNLgfpAM (A. Trkola and N. Friedrich, Institute for Medical Virology, University of Zurich, Switzerland) [32 (link)]. pCMV-VSV-G (Addgene, #12260, Teddington, UK) was used for VSV-G expression. The following plasmids were used; pME-Z1 for expression of ZIKV prME envelope proteins [8 (link)], pME-ME for expression of ME (Δpr mutant) [27 (link)], pME-E2 for expression of E2 [30 (link)], and pE41.2 for expression of E^ΔTMgp41^TMCY [30 (link)]. COS-1 cells (CVCL_0223) were provided by Friedrich Löffler Institute (Riems Greifswald, Germany). Human glioma cell lines U-87MG, U-138MG, and U-343MG were obtained from CLS Cell Lines Service (Eppelheim, Germany). All U-cells were cultured in DF medium (DMEM/10% fetal bovine serum, PAN-Biotech, Aidenbach, Germany).