Mammosphere Formation Assay Protocol

As described previously^{6 (link)}, mammosphere formation assay was performed as previously described^{6 (link)}. Single-cell suspensions of SUM149 cells (2 × 10⁴ cells/well) and BT549 cells (1 × 10⁴ cells/well), in MammoCult Human Medium Kit (StemCell Technologies, Vancouver, Canada), were seeded in 6-well ultra-low attachment plates (Corning Incorporated Costar, Corning, NY, USA). After a 7-day incubation, mammospheres were stained using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) (Sigma), and spheres greater than 80 μm in diameter were counted using the GelCount system (Oxford Optronix, UK) according to the manufacturer’s instructions.

Free full text: Click here

Gagliardi M., Pitner M.K., Park J., Xie X., Saso H., Larson R.A., Sammons R.M., Chen H., Wei C., Masuda H., Chauhan G., Kondo K., Tripathy D., Ueno N.T., Dalby K.N., Debeb B.G, & Bartholomeusz C. (2020). Differential functions of ERK1 and ERK2 in lung metastasis processes in triple-negative breast cancer. Scientific Reports, 10, 8537.

Publication 2020

MammoCult Human Medium Kit ultra-low attachment plates MTT GelCount system

Assay Bromide Cells Human Stemcell

Corresponding Organization :

Other organizations : The University of Texas MD Anderson Cancer Center, The University of Texas at Austin, Austin College

Top 5 similar protocols

Variable analysis

independent variables

Cell type (SUM149 cells and BT549 cells)

dependent variables

Mammosphere formation

control variables

Cell seeding density (2 × 10^4 cells/well for SUM149 cells and 1 × 10^4 cells/well for BT549 cells)
Culture medium (MammoCult Human Medium Kit)
Culture vessel (6-well ultra-low attachment plates)
Incubation duration (7 days)
Mammosphere size (greater than 80 μm in diameter)

controls

No positive or negative controls were explicitly mentioned.

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

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?

Revolutionizing how scientists
search and build protocols!