pHSCs and Huh7 cells were detached from two-dimensional (2D) cultures and mixed together in a ratio of 2:1 following the protocol adapted from previous studies [36 (link),46 (link)]. From this mixture, ~10,000 cells were seeded into a 96-well round bottom ultra-low attachment microplate (Corning, Corning, NY, USA; catalog #7007). Within 48 h, the co-cultured organoids will be formed and maintained in a medium containing a ratio of 1:1 of Huh7 and pHSC culture medium. The co-cultured organoids were exposed to 15 mM acetaminophen (APAP) for 24 h or 300 μM palmitic acid (PA) for 48 h to induce hepatoxicity. These organoids were then harvested for subsequent analysis.
Round bottom ultra low attachment microplate
Manufactured by Corning
Sourced in United States
The Round Bottom Ultra-Low Attachment Microplate is a laboratory equipment designed for cell culture applications. It features a round bottom well shape and a specialized surface coating that minimizes cell attachment, promoting the formation of 3D cell aggregates or spheroids. This microplate is suitable for a range of cell-based assays and experiments that require the maintenance of a suspension culture environment.
Automatically generated - may contain errors
Lab products found in correlation
Columbus software, by PerkinElmer (1 mentions)
Hoechst 33342, by Merck Group (1 mentions)
Tcf lef luciferase reporter plasmid, by Promega (1 mentions)
Fetal bovine serum (fbs), by Innoprot (1 mentions)
Silibinin, by Merck Group (1 mentions)
Ly294002, by Merck Group (1 mentions)
Epidermal growth factor (egf), by Thermo Fisher Scientific (1 mentions)
Basic fibroblast growth factor (bfgf), by Thermo Fisher Scientific (1 mentions)
Dmem f12, by Thermo Fisher Scientific (1 mentions)
Insulin, by Merck Group (1 mentions)
Mini shaker, by Avantor (1 mentions)
Pmg0034, by Thermo Fisher Scientific (1 mentions)
Phc9394, by Thermo Fisher Scientific (1 mentions)
L ascorbic acid phosphate, by Fujifilm (1 mentions)
24 well ultra low attachment plate, by Corning (1 mentions)
6 protocols using round bottom ultra low attachment microplate
1
Hepatotoxicity Evaluation in Organoid Co-culture
2
Spheroid Culture and Compound Screening
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HT-1080 cells were seeded into a 96-well Round Bottom Ultra Low Attachment Microplate (Corning costar 7007) with a density of 2000 cells per well. After 48 h of growth, spheroids were treated with RSL3 and Ferrostatin-1 or Turofexorate/Fexaramine and incubated for additional 48 h. Staining was performed by adding Hoechst 33342 (Sigma) in a 1:10,000 dilution directly into the wells and incubating for 1 h. Images were taken with an Operetta high-content system and analyzed with Columbus software (PerkinElmer). For analysis, spheroids were detected as “Image region” and morphology properties were calculated (e.g., roundness).
3
3D Spheroid Culture of Hair Follicle DP Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The human hair follicle DP cells were cultured in 5% CO 2 at 37°C in mesenchymal stem cell medium (MSCM; Innoprot) supplemented with 5% (v/v) fetal bovine serum (FBS) according to the instructions provided by Innoprot (Bizakaia, Spain). TCF/ LEF luciferase reporter plasmids were obtained from Promega (USA). A selective phosphatidylinositol 3-kinase (PI3K) inhibitor, LY294002, and silibinin were purchased from Sigma-Aldrich (USA).
Three-dimensional (3D) culture of DP cells. Three-dimensional (3D) culture of DP cells was performed as previously described [19] . Briefly, cells were maintained in 2D culture as described above. To obtain one spherical structure of the cells, cultured human DP cells (5 × 10 4 cells/well) were seeded into a 96-well, clear, round-bottom ultra-low attachment microplate (Corning Incorporated). After the formation of the unified spheres, the spheres were treated with silibinin for 48 h. Finally, the diameters of the spheres were measured by phase-contrast microscopy.
Three-dimensional (3D) culture of DP cells. Three-dimensional (3D) culture of DP cells was performed as previously described [19] . Briefly, cells were maintained in 2D culture as described above. To obtain one spherical structure of the cells, cultured human DP cells (5 × 10 4 cells/well) were seeded into a 96-well, clear, round-bottom ultra-low attachment microplate (Corning Incorporated). After the formation of the unified spheres, the spheres were treated with silibinin for 48 h. Finally, the diameters of the spheres were measured by phase-contrast microscopy.
4
Generation and Drug Testing of Tumor Spheroids
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To generate tumor spheroids, cells suspended in complete media were seeded at a density of 6 × 103 cells/well, depending on the proportions of cancer and stromal cells in 384- or 96-well round bottom ultra-low attachment microplates (Corning Life Sciences, Amsterdam, Netherlands). The plates were incubated for 3 days at 37 °C in a humidified atmosphere of 5% CO2. After 3 days, 10 µM of cytotoxic or anticancer drugs were added and incubated for an additional 7 days.
5
Huh7.5-RFP Spheroid Drug Screening
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Huh7.5-RFP cells were seeded in 96-well round bottom ultra-low attachment microplates (Corning) at a density of 1 cell/well in LCSC media. LCSC media is composed of DMEM/F12 (10565-018, Gibco) added with 1x B27 (Invitrogen), 20 ng/ml basic fibroblast growth factor (bFGF; Invitrogen), 20 ng/ml epidermal growth factor (EGF, Invitrogen), 25 µg/ml insulin (Sigma-Aldrich) (LCSC media) and cultivated at humidified 37 °C incubator under 5% CO2 for 5 days. After 5 days, homogenous and singular spheroid was transferred to new 96-well round bottom ULA microplates and treated with 4 hit drugs, sorafenib (positive control), and 0.5% DMSO (negative control) for 7 days. Bright-filed images were obtained everyday with HCS System.
6
Gastruloid Generation from mESCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Gastruloids were generated as previously described (Baillie-Johnson et al., 2015; (link)Rossi et al., 2020; (link)van den Brink et al., 2014) (link). Briefly, 300-700 mESCs were aggregated in 40μl N2B27 in 96-well Clear Round Bottom Ultra-Low Attachment Microplates (7007, Corning). After 48h, 150μl per well of 3μM Chi in N2B27 were added. At 72h, 150μl of medium were removed and substituted with 150μl of fresh N2B27. From 96h, the medium was changed to N2B27+++, which contains 30ng ml -1 bFGF (PMG0034, Gibco), 5ng ml -1 VEGF 165 (PHC9394, Gibco) and 0.5mM L-ascorbic acid phosphate (013-12061, Wako). From 120h on, half of the medium was changed daily. From 144h, N2B27 was used for daily medium changes. For immunofluorescence analysis, gastruloids at 96h were transferred in Ultra-Low Attachment 24-Well Plates (3473, Corning) with 100μl of medium, plus 700μl of fresh N2B27+++, and cultured on an orbital shaker placed at 37°C, 5% CO2 at 100rpm (VWR mini shaker), with the same culture schedule.
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!