Collagen coating of 60 mm petri dishes (BD Biosciences, Heidelberg, Germany) was done as previously described with minor modifications [31 (link)]. The pH of the collagen was adjusted to 7.4 using a NaHCO3 solution and 1 ml of a 0.5 mg/ml collagen solution was applied to coat the 60 mm petri dishes (Greiner, Frickenhausen, Germany). The dishes were prepared in advance of cell isolations. Following seeding of 2 x 106 hepatocytes per dish the cells were allowed to attach. After 24 h in culture, the medium was removed along with non-adherent cells. A second layer of collagen was pipetted on top of the cells. After gelation of the second layer culture media [2 ml of William’s E supplemented with 5% fetal calf serum (FCS; Biochrom), 9.6 μg/ml prednisolone, 0014 μg/ml glucagon (Novo, Germany), 0,16 U/ml insulin (Hoechst, Germany), 200 U/ml penicillin, and 200 U/ml streptomycin (GIBCO, Germany)] was added and changed daily. The cells were cultured at 37°C and an atmosphere of 5% CO2 and were inspected by phase contrast microscopy every 24h.
60 mm petri dishes
Manufactured by BD
Sourced in Germany, United States
The 60 mm Petri dishes are laboratory equipment used for the cultivation and containment of samples or specimens. They provide a sterile, controlled environment for various scientific applications.
Automatically generated - may contain errors
Lab products found in correlation
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Fetal calf serum (fcs), by Harvard Bioscience (1 mentions)
60 mm petri dishes, by Greiner (1 mentions)
Inverted microscope, by Nikon (1 mentions)
Bacto agar, by BD (1 mentions)
G1 plus medium, by Vitrolife (1 mentions)
Dulbecco s minimum essential medium, by Merck Group (1 mentions)
H9 hnscs, by Thermo Fisher Scientific (1 mentions)
Ham s f 12 nutrient mixture, by Merck Group (1 mentions)
6 well plate, by BD (1 mentions)
6 protocols using 60 mm petri dishes
1
Collagen-coated Hepatocyte Culture Protocol
2
Mammosphere colony formation assay
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Mammospheres were resuspended in 0.5% agar (Bacto-Agar, Difco Laboratories) and layered on a preformed 0.8% agar layer using 60 mm Petri dishes (BD). Colonies were counted under an inverted microscope (Nikon, Milan, Italy) and then photographed.
3
Colony Forming Assay for Cell Survival
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Cell survival was determined using a conventional colony forming assay. Briefly, the cells were collected by trypsinization and resuspended in RPMI-1640 medium containing 10% FBS immediately after exposure. The cell concentration was determined using a model Z1 cell Coulter counter with a 100 µm aperture tube. The cells were diluted with medium and seeded in 60 mm Petri dishes (Falcon, Germantown, Washington, DC) to obtain 10 to 100 colonies per dish. The dishes were incubated for the appropriate days corresponding to the cell lines, then fixed with 10% formaldehyde for 10 minutes and stained with crystal violet to visualize the colonies. Colonies containing more than 50 cells were counted as survivors. Three parallel dishes were scored each time for every dose.
4
Vitrification and Survival of Oocytes and Zygotes
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The survival of the oocytes and zygotes was assessed two hours after completing the warming and rehydration process: those whose morphological appearance remained similar to fresh after vitrification were considered positive for survival12 (link). As an additional viability indicator, post-warming mitosis resumption and cleavage was assessed after 24 hours of culture. Results were compared with a control group of 27 abnormal zygotes not subjected to vitrification. Embryo culture was carried out in pre-equilibrated embryotested 60 mm petri dishes (Falcon, Corning, USA), containing 30 µl drops of G1-Plus medium (Vitrolife, Sweden) with a mineral oil overlay at 37 °C and 6% CO2.
5
Culturing human neural stem cells
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The hNSCs derived from H9 hNSCs were purchased from Gibco, Japan. The hNSCs were cultured in 60 mm Petri dishes (BD Falcon) or in 6-well plates (BD Falcon) with a growth medium Knock OutTM:1:1 (v/v) mixture of Dulbecco’s minimum essential medium (Sigma, U.S.A.) and Ham’s F-12 nutrient mixture (Sigma, U.S.A.) supplemented with 2% StemPro neural supplement, 20 ng/mL of fibroblast growth factor (FGF) basic recombinant human, 20 ng/mL epidermal growth factor (EGF) recombinant human and 2 mM GlutaMAX-I supplement. For adhesion, 60-mm Petri dish or 6-well plate coated with CELLStart were used. The hNSCs were cultured at 37ºC in a 95% air / 5% CO2 humidified incubator. The medium was changed every two days to keep cells undifferentiated.
6
Isolation of U87 Cell-Derived Exosomes
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U87 cells were cultured in six 60 mm Petri dishes (falcon) with FBS-originated-exosome-free media (as instructed in the protocol by Théry; FBS was ultracentrifuged at 100 000 g for 2 hours at 4°C, then filtered with a 0.22 μm sterile filter) for 48 hours; after 48 hours, the media containing U87 exosomes was isolated. Total cell count was 2×107 and 24 mL of U87-exosome-containing media was obtained. For the following isolation methods, the same batch of media was used.
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