Human bone marrow mesenchymal stem cell line (BMSC) was purchased from AddexBio Technologies, CA, USA. Upon receiving the cells in liquid nitrogen, they were immediately washed with phosphate buffered saline (PBS, Gibco, USA) to ensure that the cells were free from the cryoprotectant Dimethyl Sulfoxide (DMSO). The washed cells were then cultured in mesenchymal stem cell growth media (MSCM, AddexBio Technologies, CA, USA) supplemented with 10% Fetal Bovine Serum (FBS, Himedia, USA) and 1% penicillin streptomycin antibiotic solution (Himedia, USA). After the addition of MSCM, the cells were kept in an incubator supplied at 37°C temperature and 5% CO2. The cell culture medium was replaced with fresh medium at an interval of three days, and their confluency appraised. Once the cells achieved 90% confluency, the proteolytic enzyme trypsin was added to dislodge the adherent cells from the culture flask. Trypsinization was done using trypsin/EDTA solution (Himedia, USA). The detached cells were sub-cultured for further growth in MSCM.
Penicillin streptomycin antibiotic solution
Manufactured by HiMedia
Sourced in India
Penicillin-streptomycin antibiotic solution is a combination of two commonly used antibiotics, penicillin and streptomycin. The solution is designed for laboratory use and serves as a broad-spectrum antimicrobial agent to prevent bacterial contamination in cell culture and other in vitro applications.
Automatically generated - may contain errors
Lab products found in correlation
Trypsin edta solution, by HiMedia (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by HiMedia (1 mentions)
Dulbecco modified eagle medium (dmem), by HiMedia (1 mentions)
Trypsin, by HiMedia (1 mentions)
Ethylene diamine tetraacetic acid (edta), by HiMedia (1 mentions)
Acridine orange, by Merck Group (1 mentions)
Bisbenzimide hoechst 33342, by Merck Group (1 mentions)
5 bromo 4 chloro 3 indolyl phosphate bcip nitroblue tetrazolium nbt, by Merck Group (1 mentions)
Histopaque, by Merck Group (1 mentions)
Paraformaldehyde (pfa), by Merck Group (1 mentions)
Nabh4, by Merck Group (1 mentions)
Propidium iodide, by Merck Group (1 mentions)
Silver nitrate agno3, by Merck Group (1 mentions)
Dulbecco s modified eagle s medium dmem, by HiMedia (1 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by HiMedia (1 mentions)
Polyethylene glycol peg, by Merck Group (1 mentions)
3 protocols using penicillin streptomycin antibiotic solution
1
Propagation of Human Bone Marrow Mesenchymal Stem Cells
2
Cytotoxicity Evaluation of Hydrogel Films
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Cytotoxicity assay of the prepared hydrogels was performed by MTT colorimetric technique using 9 mm diameter discs of the hydrogel films. Prior to the assay, the hydrogels were sterilized at 15 lb/in2 steam pressure, 121 °C for 1 h. The cytotoxicity was evaluated by using L929NCCS rat fibroblast cell line. The cell growth was performed on a 24-well tissue culture plate in a controlled atmosphere (5% CO2 at 37 °C) using a cell culture medium of Dulbecco's Modified Eagle's Medium (DMEM, Hi-Media, India) supplemented with 10% Fetal Bovine Serum (FBS, Hi-Media, India) and penicillin–streptomycin antibiotic solution (Hi-Media, India). 90% of confluent monolayers of cultured cells were harvested by Trypsinization (0.25% Trypsin and 0.02% EDTA, Hi-Media, India) and 1 mL of 1 × 105cells/mL were seeded in each well. The culture plate was then incubated for 48 h in the CO2 incubator at 37 °C. The cytotoxicity analysis was performed as per standard protocol.
3
Phytochemical Extraction and Bioactivity Evaluation
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Mentha arvensis leaves were collected from the state of Assam, India. Silver nitrate (AgNO3), poly(ethylene glycol) (PEG) and 5-bromo-4-chloro-3-indolyl phosphate (BCIP)/nitro blue tetrazolium (NBT) were purchased from Merck (India). Dulbecco’s Modified Eagle’s Medium (DMEM), Roswell Park Memorial Institute (RPMI)-1640 culture media, fetal bovine serum (FBS), penicillin–streptomycin antibiotic solution, phytohemagglutinin (PHA), 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) and radio-immunoprecipitation assay (RIPA) buffer were purchased from HiMedia (Mumbai, India). Heparin, ethidium bromide (EtBr) and trypan blue were purchased from SRL (India); poly-l-lysine, Histopaque, bisbenzimide (Hoechst 33342), paraformaldehyde, acridine orange, propidium iodide (PI), RNase, NaBH4 and anti-human primary antibodies used were procured from Sigma-Aldrich (St Louis, MO, USA). ALP-linked goat anti-rabbit secondary antibodies were purchased from Abcam (UK).
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