We examined the morphological modifications on treated cells, compared to untreated control cells. Cells were seeded in 6 well plates using the RPMI medium. After incubation in a humidified 37 °C, 5% CO2 incubator (Series II Water Jacket supplied by Thermo Scientific) overnight, cells were then treated with respective clam extracts (600 μg/ml) and incubated for 72 h. Cell morphology was observed under an inverted microscope (Zeiss Primotech microscope), at 40 x magnification.
Series 2 water jacket
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Series II Water Jacket is a laboratory equipment piece designed for cell culture applications. It maintains a stable and consistent temperature environment within the incubator chamber to support optimal cell growth and proliferation.
Automatically generated - may contain errors
Lab products found in correlation
Primotech microscope, by Zeiss (1 mentions)
Phorbol myristate acetate (pma), by Abcam (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Dulbecco phosphate buffered saline (dpbs), by BasalMedia (1 mentions)
Stempro accutase cell dissociation reagent, by Thermo Fisher Scientific (1 mentions)
Parafilm, by Thermo Fisher Scientific (1 mentions)
5 protocols using series 2 water jacket
1
Morphological Changes in Treated Cells
2
Cytotoxic Potential of NZ Surf Clam Extracts
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The cytotoxic effect of NZ surf clam extracts on three cancer cell lines was measured employing the MTT assay. The cells were seeded in a 96-well plate at a concentration of 1 × 105 cells mL−1 using the RPMI medium supplemented with 1 % Penicillin-Streptomycin, 1 % l -glutamine and 10 % fetal bovine serum. After incubation in a humidified 37 °C, 5% CO2 incubator (Series II Water Jacket supplied by Thermo Scientific) overnight, the cells were treated by NZ surf clam extracts at a concentration range from 25 to 1000 μg/mL. The cells were further incubated for an additional 24, 48, and 72 h independently at 37 °C. After incubation, MTT stock solution was then added to each well and incubated for a further 4 h. The formazan crystals in each well were dissolved in 100 μL of DMSO. The amount of purple formazan was determined by measuring the absorbance at 540 nm.
3
Culturing Diverse Lung Cell Lines
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A549 (human lung adenocarcinoma cell line, ATCC, USA), HLF (human lung fibroblast cell line, ATCC) and 293 T (human kidney epithelial cell line, ATCC) cells were grown in Dulbecco’s modified Eagle’s medium (DMEM, Gibco, USA) containing 10% foetal bovine serum (FBS, ExCell Bio, Shanghai, China) and 1% penicillin‒streptomycin solution. THP-1 cells (human monocyte cell line, ATCC) and MLE12 cells (mouse lung epithelial cell line, ATCC) were grown in RPMI-1640 (Gibco, USA) supplemented with 10% FBS and 1% penicillin‒streptomycin solution. All of the above cells were cultured at 37 °C in a humidified incubator with 5% CO2 (Thermo Scientific, Series II Water Jacket). THP-1 cells were differentiated into macrophages using 200 μg/mL phorbol 12-myristate 13-acetate (PMA, Abcam, USA).
4
Isolation of Lung Macrophages from Human and Mouse
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Human lung macrophages and mouse lung macrophages were isolated as previously described [22 , 23 (link)]. Briefly, in a cell culture dish containing Dulbecco’s phosphate buffered saline (DPBS, BasalMedia, Shanghai, China), human lung tissues were cut into small pieces, which were gently shaken and then removed. The liquid in the dish was a cell suspension containing human lung macrophages. Mouse lung macrophages were obtained from bronchoalveolar lavage fluid (BALF). For a single sample, the cell suspension was collected into a centrifuge tube and centrifuged at 700×g and 4 °C for 5 min. The supernatant was discarded, and the cell pellet was resuspended in erythrocyte lysate (Boster, Wuhan, China) at room temperature (RT) for 5 min and then centrifuged at 700×g and 4 °C for 5 min. The supernatant was then discarded, and the cell pellet was resuspended in RPMI-1640 and added to a cell culture plate. The cells were allowed to adhere to the plate for 1–2 h at 37 °C in a humidified incubator with 5% CO2 (Series II Water Jacket, Thermo Scientific, USA). Nonadherent cells were removed by gently washing three times with warm PBS. Other irrelevant adherent cells, such as lung epithelia, were removed by digestion with Accutase (StemPro™ Accutase™ Cell Dissociation Reagent, Gibco). Eventually, greater than 90% of the cultured cells were macrophages.
5
Sub-lethal Thermal Stress Responses
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SMMC-7721, HepG2, and MHCC97-H cells were sub-lethally heated (50 °C) for 10 min. Heat treatments were carried out by sealing the culture bottle with Parafilm, and submerging the plates in a water bath (HH · W21 · 600S, Shanghai, China) set at 50 °C and returned to the incubation chamber (Series II Water Jacket, Thermo-Scientific, Waltham, MA) for 24 h at 37 °C. Three independent experiments were performed.
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