Minimum Essential Medium Eagle - Earle’s (EMEM medium), Fetal Bovine Serum (FBS), Penicillin-Streptomycin-Nystatin Solution, and Dulbecco’s Phosphate Buffered Saline (DPBS) were obtained from Biological Industries (Bet-Haemek, IL). Opti-MEM medium was obtained from Gibco-ThermoFisher (Grand Island, NY). TransIT-X2 Mirus reagent was also used (MC-MIR-6003; Madison, WI).
Penicillin streptomycin nystatin solution
Manufactured by Sartorius
Sourced in Israel, United States
Penicillin-Streptomycin-Nystatin Solution is a sterile, liquid media supplement composed of the antibiotics penicillin and streptomycin, as well as the antifungal agent nystatin. It is commonly used to prevent bacterial and fungal contamination in cell culture applications.
Automatically generated - may contain errors
Lab products found in correlation
Opti mem medium, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Sartorius (1 mentions)
Dulbecco s phosphate buffered saline, by Sartorius (1 mentions)
Facscanto 2, by BD (1 mentions)
Lsm800 confocal microscope, by Zeiss (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Corning (1 mentions)
Jetprime reagent, by Polyplus Transfection (1 mentions)
L alanyl l glutamine, by Sartorius (1 mentions)
Dulbecco s modified eagle medium dmem, by Sartorius (1 mentions)
Puromycin, by InvivoGen (1 mentions)
Hamec, by ScienCell (1 mentions)
Glutamax, by Thermo Fisher Scientific (1 mentions)
Mitomycin, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by ScienCell (1 mentions)
Endothelial cell growth supplement (ecgs), by ScienCell (1 mentions)
Endothelial cell medium (ecm), by ScienCell (1 mentions)
Pen strep, by Sartorius (1 mentions)
Dulbecco s modified eagle medium dmem, by Thermo Fisher Scientific (1 mentions)
Low glucose dmem, by Thermo Fisher Scientific (1 mentions)
4 protocols using penicillin streptomycin nystatin solution
1
Cell Culture Protocol for Transfection
2
Uptake and Cytotoxicity of Gold Nanoparticles
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PC-3 cells were maintained in RPMI-1640 medium (Corning Incorporated) supplemented with 10% FBS (heat inactivated; Thermo Fisher Scientific, Waltham, MA, USA) and 1% penicillin-streptomycin nystatin solution (Biological Industries). Cells (5×104 per well) were seeded in 24-well culture plates for 24 hours. Cells were then treated with Au800-CTAB-PAA-PEG or Au800-CTAB-PAA-PEG-AA (5 µg/mL) and incubated for 8 hours under normal growth conditions. Following incubation, cells were washed twice with PBS. Fresh growth medium was added to the cells prior to confocal microscopic analysis using an LSM 800 Zeiss confocal microscope (40×).
In addition, cells were also treated with Au800-CTAB-PAA-PEG or Au800-CTAB-PAA-PEG-AA (5 µg/mL) containing EPI (MR =5) and incubated for 4 hours under normal growth conditions. After this, cells were washed twice with PBS and trypsinized. After 1,000 rpm centrifugation for 5 minutes, the supernatant was discarded and cells were resuspended in 1,000 µL of ice-cold PBS. About 10,000 cells were measured to determine EPI-positive cells (%) for each sample using flow cytometry (Becton Dickinson FACS-Canto II).
In addition, cells were also treated with Au800-CTAB-PAA-PEG or Au800-CTAB-PAA-PEG-AA (5 µg/mL) containing EPI (MR =5) and incubated for 4 hours under normal growth conditions. After this, cells were washed twice with PBS and trypsinized. After 1,000 rpm centrifugation for 5 minutes, the supernatant was discarded and cells were resuspended in 1,000 µL of ice-cold PBS. About 10,000 cells were measured to determine EPI-positive cells (%) for each sample using flow cytometry (Becton Dickinson FACS-Canto II).
3
ADAR Enzyme Expression and Selection
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Cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM) (Biological Industries, 01-055-1A) supplemented with L-Alanyl-L-Glutamine (Biological Industries, 03-020-1B), Foetal Bovine Serum (FBS) European Grade Heat Inactivated (Biological Industries, 04-127-1A) and Penicillin-Streptomycin-Nystatin Solution (Biological Industries, 03-032-1B). mdADAR1, hbADAR2 and hADAR2 were synthesized by Twist Bioscience with an N’ Flag (DYKDDDDK) and cloned into pTwist-CMV promoter-Puromycin selection plasmids. Codons were optimized using Twist codon optimization tool. Transfection was carried out using jetPRIME reagent (Polyplus Transfection). Puromycin (InvivoGen ant-pr-1) was used for selection.
4
3D Vascularized Tissue Constructs
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Human adipose microvascular endothelial cells (HAMECs; ScienceCell), lentivirally transduced with ZsGreen fluorescent protein, were grown in endothelial cell medium (ScienceCell), supplemented with 5% FBS (ScienceCell), and endothelial cell growth supplement (ScienceCell), and used for five to nine passages. Neonatal normal human dermal fibroblasts (HNDFs) (Lonza Walkersville Inc.) were grown in Dulbecco’s modified Eagle medium (DMEM) (Gibco), supplemented with 10% FBS (HyClone), 1% non-essential amino acids (NEAAs), 0.2% β-mercaptoethanol (Sigma-Aldrich), and 1% penicillin–streptomycin solution (PEN STREP) (Biological Industries). Dental pulp stem cells (DPSCs) (Lonza) were cultured in low glucose DMEM (Gibco), supplemented with 10% FBS (HyClone), 1% NEAAs, 1% GlutaMAX (Gibco), and 1% penicillin–streptomycin-nystatin solution (Biological Industries). Three dimensional, vascularized constructs were obtained by coseeding endothelial cells (3 × 105 cells) and DPSCs (9 × 105) or fibroblasts (0.6 × 105 cells) in 20 µl medium on the gelfoam or in 7 µl fibrin on the PLLA/PLGA scaffolds followed by incubation of 30 min, before addition of medium. For the mitomycin experiments, 5 ml mitomycin (Sigma-Aldrich) was applied to the fibroblast cells 2 h before seeding, and washed twice with PBS.
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