Three-day-old ICR mice were used for primary microglial culture. Mice were anesthetized in cold ice and decapitated. After removal of skull and meninges, cortex and hippocampus were extracted and dipped in HBSS in 15 ml conical tube. Tissue was mechanically dissociated through pipetting. Then, trypsin was added to tissue in 1:1 ratio (4 ml each) with HBSS and incubated in 37 °C water bath for 15 min. During incubation, conical tube was inverted every 3 min. Then, 2 ml of FBS was added and centrifugated at 700 rpm for 15 min. Supernatants were removed, and tissue was dissociated and cultured with primary culture medium (FBS 5 ml, Horse serum 5 ml, penicillin-streptomycin 500 μl, glutaMAX 500 μl, 25 ng m-CSF, DMEM F12 in 50 ml) in cell culture flask (SPL life science) with 16 ml of media in each flask. Media was changed 2~3 days after the incubation. Then, half of the media was changed every 2 days until harvest. Whole media was changed a day before harvest and primary microglia was harvested by shaking culture flasks in 260 rpm for 3h. Then, media was collected and centrifuged at 700 rpm for 15 min. Then, cell pellets were harvested and cultured on glass chip on 12~24 well plate and used for analysis [46 (link)].
Cell culture flask
Manufactured by SPL Life Sciences
Sourced in Germany
The cell culture flask is a laboratory equipment used for the in vitro cultivation of cells. It provides a controlled and sterile environment for cells to grow and proliferate. The flask is made of transparent, gas-permeable material, typically polystyrene or borosilicate glass, and is equipped with a screw-cap or snap-on lid to maintain the culture conditions.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Hela cell, by Thermo Fisher Scientific (1 mentions)
Dmem medium, by Thermo Fisher Scientific (1 mentions)
Hela cells, by SPL Life Sciences (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dmem medium, by Biosera (1 mentions)
Inverted microscope, by Labomed (1 mentions)
Antibiotic antimycotic solution, by Biosera (1 mentions)
Cd8 t cell isolation kit, by Miltenyi Biotec (1 mentions)
Taq dna polymerase master mix red, by Ampliqon (1 mentions)
Minimacs, by Miltenyi Biotec (1 mentions)
Annexin 5 pe apoptosis detection kit, by Thermo Fisher Scientific (1 mentions)
3 4 5 dimethyl 2 thiazolyl 2 5 diphenyl 2h tetrazolium bromide mtt, by Merck Group (1 mentions)
Trypan blue solution 0.4, by Merck Group (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Reverse transcriptase reagent kit, by Thermo Fisher Scientific (1 mentions)
25 cm2 cell culture flasks, by Thermo Fisher Scientific (1 mentions)
Complete rpmi 1640 medium, by PAN Biotech (1 mentions)
6 protocols using cell culture flask
1
Primary Microglial Culture from Mouse Cortex and Hippocampus
2
Evaluating ZnO Nanoparticle Cytotoxicity in HeLa Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HeLa cells were purchased from the Korea Biological Resource Center (Daejeon, Korea). HeLa cells were cultured in fresh DMEM medium (Life Technologies Corp) including 10% FBS and 1% penicillin-streptomycin (15140-122, Life Technologies Corp). Cells suspensions, prepared at a concentration of 104–105 cells/mL were loaded in the 96-well plate. Freshly isolated HeLa cells were cultured in a plastic Cell culture flask (SPL Lifesciences Co. Ltd., Seoul, Korea). After seeding, cells were maintained at 37°C in a 5% CO2 incubator for 36 hours. Then, the HeLa cells were treated with 0, 10, 20, 30, 40, 50, and 60 mg/L ZnO, ZnO(PAA), or ZnO(PLL) in DMEM media without FBS or with FBS for 24 hours.
3
Isolation and Characterization of Human Dermal Fibroblasts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human dermal fibroblast cells were purchased from Bon Yakhte Corp. (Tehran, Iran). First, the cells were examined for CD34, CD45, CD73, CD90 and CD105 biomarkers expressions by flowcytometry. 4 mL of high glucose content (4.5 g/L) DMEM medium (Biosera, France) containing 10% FBS (Gibco, USA) and 1% antibiotic-antimycotic solution (Biosera, France) were used for cell culture. The culture medium in the Falcon was filled with the cell suspension, which was then carefully pipetted into the container. The cell-containing tube was then spun at 230 g for 5 minutes (VISION SCIENTIFIC, South Korea). After removing the supernatant, the cell sediment was suspended in 1 mL of culture medium, and transferred to a 25 cm cell culture flask (SPL, South Korea) containing 4 mL of complete culture medium. After observing the cells inside the flask under an inverted microscope (LaboMed, USA), the flask was transferred to an incubator (MEMMERT, Italy) with a temperature of 37 °C, 5% CO2, and 95% relative humidity (RH).
The percentage of viable cells was determined by cell staining with trypan blue. For this purpose, at first 1 mL cell suspension was prepared, and 20 µL of trypan blue 0.25% was added to cell suspension and poured into a well of a 96-well plate. About 10 µL of the mixture was placed on Neubauer slide and transferred under the microscope.
The percentage of viable cells was determined by cell staining with trypan blue. For this purpose, at first 1 mL cell suspension was prepared, and 20 µL of trypan blue 0.25% was added to cell suspension and poured into a well of a 96-well plate. About 10 µL of the mixture was placed on Neubauer slide and transferred under the microscope.
4
Isolation and Analysis of T-cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All cell culture media and components were provided from Gibco, Life Technologies (Paisley, UK). qPCR SYBER Green master mix was purchased from BioFACT (Daejeon, Korea), and Taq DNA Polymerase Master Mix RED was obtained from Ampliqon. Well plates, cell culture flasks, and pipettes were purchased from SPL Life Sciences (Pocheon, South Korea).Anti-LC3 antibody (MAP LC3α/β SC-398822 F2918), Annexin V Apoptosis Detection Kit PE (eBioscience), CD8+ T cell isolation kit, MiniMACS (Miltenyi Biotec Inc cat NO: 130-096-543), AO (Invitrogen, USA).
5
Pistachio Pollens and Stem Cell Interactions
Check if the same lab product or an alternative is used in the 5 most similar protocols
The mature PGs of Pistachio (Pistacia vera L.) were collected from male flowers in early spring in Qazi Jahan Pistachio gardens, Iran. The hAD-MSCs were obtained from the Iranian Biological Resource Center (Tehran, Iran). Trypan blue solution (0.4%) and 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) were purchased from Sigma-Aldrich (Poole, UK). TRIzol reagent and reverse transcriptase reagent kit were bought from Thermo Fisher Scientific (Waltham, MA, USA). Maxima SYBER Green qPCR master mix was obtained from BioFACT (Daejeon, Korea). Cell culture flasks, well plates, and pipettes were purchased from SPL Life Sciences (Pocheon, South Korea). All cell culture media and components were provided from Gibco, Life Technologies (Paisley, UK). The oligonucleotide primer sequences were received from Takapo Zist Company (Tehran, Iran). Other chemicals were supplied from Merck (Kenilworth, NJ, USA).
+ Expand
The mature PGs of Pistachio (Pistacia vera L.) were collected from male flowers in early spring in Qazi Jahan Pistachio gardens, Iran. The hAD-MSCs were obtained from the Iranian Biological Resource Center (Tehran, Iran). Trypan blue solution (0.4%) and 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) were purchased from Sigma-Aldrich (Poole, UK). TRIzol reagent and reverse transcriptase reagent kit were bought from Thermo Fisher Scientific (Waltham, MA, USA). Maxima SYBER Green qPCR master mix was obtained from BioFACT (Daejeon, Korea). Cell culture flasks, well plates, and pipettes were purchased from SPL Life Sciences (Pocheon, South Korea). All cell culture media and components were provided from Gibco, Life Technologies (Paisley, UK). The oligonucleotide primer sequences were received from Takapo Zist Company (Tehran, Iran). Other chemicals were supplied from Merck (Kenilworth, NJ, USA).
6
Culturing Leishmania Parasites and Macrophages
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
L. infantum (zymodeme GH8, strain MHOM/GR/2001/GH8) and L. major (zymodeme LV39, strain MRHO/SU/59/P) parasites, causative agents of visceral and cutaneous leishmaniasis, respectively, were used in this study. Promastigotes were grown at 26 °C in complete RPMI-1640 medium (PAN-Biotech, Aidenbach, Passau, Germany) in cell culture flasks (SPL Life Sciences, Naechon-Myeon, Pocheon-si, Korea), as previously reported [48 (link)].
The immortalized macrophage cell line J774A.1 (ATCC No: TIB-67) was cultured in 25 cm2 cell culture flasks (ThermoFisher Scientific, Waltham, MA, USA), at 37 °C with a 5% CO2 environment [48 (link)].
The immortalized macrophage cell line J774A.1 (ATCC No: TIB-67) was cultured in 25 cm2 cell culture flasks (ThermoFisher Scientific, Waltham, MA, USA), at 37 °C with a 5% CO2 environment [48 (link)].
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!