Dulbecco s modified eagle s medium dmem

Rodent glioma cell line F98, the human glioma cell line U87, and the murine glioma cell line GL261 were obtained from ATCC/LGC (Wesel, Germany). The human glioma cell line U251 was provided by Yvonne Ruebner and Rainer Fietkau (Department of Radiation Therapy, Erlangen University Hospital). Primary rat astrocytes were prepared from up to 1-month-old Wistar rats. All cells were cultured under standard humidification conditions (37°C, 5% CO₂) with Dulbecco's modified Eagle's medium (DMEM; Biochrom, Berlin, Germany) supplemented by 10% fetal bovine serum (Biochrom), 1% penicillin/streptomycin (Biochrom), and 1% glutamax (Gibco/Invitrogen, Darmstadt, Germany). Cells were passaged at ∼80% confluence. Cells were scrapped of or trypsinized after the phosphate buffered saline (PBS) wash step. Cells were plated out in a culture flask after centrifugation (900 rpm for 5 min).

Poly (DL-lactide-co-glycolide) (PLGA) Resomer RG 502H and Poly(DL-lactide-co-glycolide)-co-polyethylene glycol diblock Resomer RGP d50155 were obtained from Evonik Industries AG (Darmstadt, Germany). Polyvinyl alcohol (PVA) 30,000–70,000 Da and didodecyldimethylammonium bromide (DMAB) were purchased from Sigma Aldrich (Steinheim, Germany).
For cell culture experiments Dulbecco’s modified Eagle’s medium (DMEM) and all used supplements were received from Biochrom AG (Berlin, Germany). VECTASHIELD Mounting Media with DAPI (4′,6-Diamidin-2-phenylindol) was purchased from Vector Laboratories Inc. (Burlingname, USA) and Wheat germ agglutinin (WGA) AlexaFluor 350 from Life Technologies (Carlsbad, USA). All other chemicals were obtained from Roth (Karlsruhe, Germany).

The renal cell line Vero was originally obtained from LGC Standards (Teddington, Middlesex, UK). Dulbecco’s Modified Eagle’s Medium (DMEM), l-alanine-glutamine, fetal bovine serum (FBS), penicillin/streptomycin and trypsine/EDTA were purchased from Biochrom Ltd. (Cambridge, UK). The PSA antibody and all other reagents were ordered from Sigma-Aldrich (Munich, Germany). Thirty nine whole blood samples with PSA values measured by IRMA technique in the range from zero up to 224.4 ng/mL, were collected from an equal number of patients at Army Share Fund Hospital of Athens. Prior to assay, samples were stored at −20 °C for one up to three months. All the screen printed electrodes were provided from DropSens (Llanera, Asturias, Spain).

Glomeruli isolation was carried out as described by Sharma et al. 20. Briefly, a high mid‐line incision on the abdomen exposed the abdominal cavity and total bilateral nephrectomy was performed to anesthetized animals. The kidney capsules were removed, and glomeruli were isolated following consecutive passage of the mashed cortexes through screens of 80‐ and 200‐mesh size. Glomeruli were recovered from atop the 200‐mesh screen into Dulbecco's modified Eagle's medium (DMEM; Biochrom Gmb, Berlin, Germany) supplemented with 10% foetal calf serum (FCS; Biochrom Gmb),15 mM HEPES (Biochrom Gmb), 5 mg/ml insulin‐transferrin‐sodium selenite (ITS; Sigma‐Aldrich), 50 nM dexamethasone (Sigma‐Aldrich), 2 mM glutamine (Biochrom Gmb) and 5 mM or 25 mM D‐glucose (Sigma‐Aldrich) and incubated at 37°C in a 5% (v/v) CO₂ humidified chamber for 4 or 8 days. The isolated rat glomeruli were treated with 150 nM or 300 nM paricalcitol (VDRA; Abbott, Hellas (BGP, Attika, Greece)) for 4 days in presence of normal (5 mM) or high (25 mM) glucose levels.

The C2C12BRELuc reporter cell line [41 (link)] was generated by the Inman group and used in this study. In this cell line, the luciferase (Luc) reporter is under the control of the BMP response elements (BRE) of the Id1 gene. C2C12BRELuc were cultivated in culture medium (Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% heat-inactivated fetal bovine serum (FBS) and antibiotics (100 U/mL penicillin and 100 μg/mL streptomycin, all from Biochrom AG, Berlin, Germany). C2C12BRELuc cells were passaged twice per week in a 175 or 225 cm² cell culture flask. The cultivation media was supplemented with the antibiotic G418 (0.7 mg/mL) to select for the BREluc-positive C2C12BRELuc cells in accordance with previous studies [41 (link)].
C2C1BRELuc cells were sub-cultivated at a ratio of 2.5–5 × 10⁴ cells per mL of culture medium. DMEM, antibiotics, FBS, and trypsin (Biochrom AG, Berlin, Germany) were used in cell cultivation and passaging. To ensure the viability and to calculate cell number, the metabolic activity of cells was calculated through Presto Blue^® assay (Invitrogen by Life Technologies Co., Carlsbad, CA, USA). BMP2 was purchased from Osteogenetics (Würzburg, Germany). H89 was purchased from AbCam (Cambridge, UK). Deta NONOate, SNAP, YC-1, LY83583, LDN-193189, and IBMX were purchased from Cayman Chemical (Ann Arbor, MI, USA).

Doxorubicin, superoxide dismutase (SOD), iron chloride (FeCl₃), nitro blue tetrazolium (NBT), ascorbic acid (AA), catalase (CAT) from bovine liver, acetaminophen (APAP), 2,2′-Diphenyl-1-picrylhydrazyl (DPPH), n-alkanes (C6–C30), thiobarbituric acid (TBA), trichloroacetic acid (TCA), ethylenediaminetetra acetic acid (EDTA), nicotinamide-adenine dinucleotide phosphate (NADPH), butylated hydroxytoluene, gallic acid, sodium carboxymethyl cellulose (CMC), phenazine methosulfate, Folin–Ciocalteu reagent, reduced glutathione (GSH), and 1,2-dithio-bis nitro benzoic acid (DTNB) were purchased from Sigma Co. (St. Louis, MO, USA). Dimethyl sulfoxide (DMSO) was purchased from Merck (Merck, Darmstadt, Germany). Streptomycin, Dulbecco’s modified Eagle’s medium (DMEM), and Streptomycin were purchased from Biochrom (Biochrome AG, Berlin, Germany, A 321-44). Fetal bovine serum was purchased from Sigma (Sigma, Darmstadt, Germany). Potassium phosphate monobasic, sodium pyrophosphate dibasic, sodium carbonate (Na₂CO₃), disodium hydrogen phosphate (Na₂HPO₄), hydrogen peroxide (H₂O₂), potassium ferricyanide (K₃Fe(CN)₆), sodium sulphate anhydrous (Na₂SO₄), acetic acid (ACA), and n-butanol (99.8%) were of analytical grade and purchased from Merck (Nottingham, UK).