Tumor sphere assay was performed under non-adherent and serum-free conditions as previously described as stem cell-selective conditions.25 (link),46 (link) Briefly, cells were suspended in 0.8% methylcellulose-based medium (Stem Cell Technologies, Vancouver, Canada) supplemented with 20 ng/mL epidermal growth factor (BD Biosciences, Franklin Lakes, NJ), basic fibroblast growth factor and 4 mg/mL insulin (Sigma-Aldrich, St Louis, MO) and plated at 5×103 cells in ultralow adherent 24-well plates (Corning, Corning, NY). Cells were cultured for two or three weeks and visualized under a light microscope.
Methylcellulose based medium
Manufactured by STEMCELL
Sourced in Canada
Methylcellulose-based medium is a cell culture supplement used to support the growth and differentiation of various cell types, including hematopoietic stem and progenitor cells. The medium provides a semi-solid matrix that helps maintain the undifferentiated state of cells and promotes the formation of cellular colonies or spheres.
Automatically generated - may contain errors
Lab products found in correlation
4 protocols using methylcellulose based medium
1
Tumor Sphere Assay for Stem Cells
2
Murine Hematopoietic Stem Cell Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Lin–Sca-1+CD45+CD38+c-Kit−H-2 Kb+ “mystery” population cells and Lin–Sca-1+CD45−CD38±c-Kit−H-2 Kb+ VSELs freshly sorted from murine BM were plated (104) over OP9 cells in -MEM medium supplemented with 20% FBS (Molecular Probes®, Invitrogen, Waltham, MA, USA) for 5 days and subsequently trypsinized, washed by centrifugation in α-MEM, and replated in methylcellulose-based medium (StemCell Tech, Vancouver, Canada). methylcellulose cultures were solubilized and trypsinized, and the resulting cells were washed by centrifugation in α -MEM and plated into secondary methylcellulose cultures. Cells were grown in the presence of the same growth factors and replated after 5 days into new methylcellulose cultures. The same procedure was repeated for the next 2 passages.
3
Tumor Sphere Assay for Stem Cells
4
Clonogenic Potential of Rare Hematopoietic Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
VSELs, “mystery” population cells and HSCs freshly isolated from BM (104), as well as cells harvested from OP9 cultures (primary cultures), were plated in methylcellulose-based medium (StemCell Tech, Vancouver, Canada) supplemented with murine stem cell growth factor (SCF), interleukin-3 (IL-3), granulocyte–macrophage colony-stimulating factor (GM-CSF), FLT3, thrombopoietin (TpO), erythropoietin (EpO), and insulin growth factor-2 (IGF-2). Cells were cultured for 5 days and the colonies formed were scored under an inverted microscope.
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!