Nisin Z (2.5% w/w, N5764), Pepsin (P0525000), MTT (M2003), and Cytotoxicity Detection Kit PLUS (LDH) were obtained from Sigma Aldrich (Milan, Italy). FITC Annexin-V apoptosis kit I (BD Pharmigen™, BD Biosciences, and San Jose, CA, USA, 556547) and fetal bovine serum (FBS) were purchased of Cegrogen-Biotech (Germany, A0100-3010). Pen-Strep (10,000 Unit/mL penicillin and 10,000 unit/mL streptomycin) and 0.25% trypsin-EDTA were purchased from Bioidea (Tehran, Iran). RPMI was prepared from GIBCO Laboratories (Grand Island, NY, USA, 11530586). Coumarin-6 and Oxaliplatin were kindly gifted by Professor Valizadeh’s laboratory (TUOMS, Tabriz, Iran). All other chemicals were of analytical grade.
Pen strep
Manufactured by Bioidea
Pen/strep is a common antibiotic mixture used in cell culture and molecular biology applications. It contains the antibiotics penicillin and streptomycin, which together provide broad-spectrum antimicrobial activity to prevent bacterial contamination in various experimental procedures.
Automatically generated - may contain errors
Lab products found in correlation
Methyl thiazolyl tetrazolium (mtt), by Merck Group (2 mentions)
Dulbecco modified eagle medium (dmem), by Bioidea (2 mentions)
Phosphate buffered saline (pbs), by Bioidea (2 mentions)
Fetal bovine serum (fbs), by Bioidea (2 mentions)
Cytotoxicity detection kitplus ldh, by Merck Group (1 mentions)
Nisin z, by Merck Group (1 mentions)
Fitc annexin 5 apoptosis kit 1, by BD (1 mentions)
Trypsin edta, by Bioidea (1 mentions)
Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (1 mentions)
Pepsin, by Merck Group (1 mentions)
ε caprolactone ε cl, by Merck Group (1 mentions)
Glutaraldehyde 25 aqueous solution, by Merck Group (1 mentions)
Poly ε caprolactone, by Merck Group (1 mentions)
Collagen type 1, by Merck Group (1 mentions)
Trypsin, by Bioidea (1 mentions)
4 protocols using pen strep
1
Cytotoxicity and Apoptosis Evaluation
2
Collagen-PCL Scaffold Fabrication
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Collagen type I, stannous 2-ethyl hexanoate (stannous octoate, Sn(Oct)2) and MTT were purchased from Sigma-Aldrich (Chemical Co., St. Louis, MO). Glutaraldehyde (25% aqueous solution), and all the solvents were purchased from Merck Chemical Co. (Darmstadt, Germany). Poly-ε-caprolactone (PCL) was synthesized from ε-caprolactone (ε-CL) monomers purchased from Merck Chemical Co. (Darmstadt, Germany). Also, fetal bovine serum (FBS), DMEM, Pen/Strep, and trypsin were purchased from Bio idea. Polymer specifications are presented in Table 1 .
3
MG-63 Cell Culture and SEM Analysis
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MG-63 cell line cells were cultured in a Dulbecco's modified Eagle's medium (DMEM, Bio-Idea, Iran) containing 10% fetal bovine serum (FBS, Bio-Idea, Iran) and 1% Penicillin/streptomycin (Pen/strep, Bio-Idea, Iran) for several passages to reach a stable phenotype. About 10,000 cells were seeded on each sample and were incubated in 37°C and 5% CO2 atmosphere. After 1 and 7 days, samples were washed with phosphate-buffered saline (PBS, Bio-Idea, Iran) to eliminate unattached cells and fixed in 2% glutaraldehyde for 30 min at room temperature. After several dehydrations in ethanol (30 min in 50%, 70%, 80%, 90%, and 100% ethanol, subsequently), SEM (Philips XL30, The Netherlands) was used to study cells morphology on the surface.
4
Isolation and Propagation of Viral Cultures
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The CSF sample was inoculated and grown on a human rhabdomyosarcoma (RD) cell line at 37 °C in a 5% CO 2 atmosphere for 5-6 days. The cells were observed for signs of cytopathic effects (CPEs) for 24 h. The medium in each well was then decanted, the wells were washed twice with phosphate-buffered saline (PBS, Bio-Idea, Iran), and then the medium was replaced with fresh 1x Dulbecco's Modified Eagle Medium (DMEM; Bio-Idea, Iran) supplemented with 10% heat inactivated fetal bovine serum (FBS; Bio-Idea, Iran) and 1% penicillin (100 U/mL)/streptomycin (100 mg/ml) (Pen/strep, Bio-Idea, Iran). Subsequently, when the CPEs were observed, suspensions of pure-culture virus were prepared by multiple freezing (-20 °C) and thawing (37 °C) cycles and a final vortexing.
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