The murine recombinant cytokines SCF, TPO, IL-3, IL-6 and FLT3L were obtained from PeproTech MethoCult M3234 Methylcellulose medium was obtained from Stem Cell Technologies. The following antibodies for flow cytometry were obtained from eBioscience: CD45.1 (A20); CD45.2 (104); Sca-1 (D7); c-Kit (2B8); CD16 (93). CD34 (RAM34) and FLT3 (A2F10.1) antibodies were obtained from BD biosciences. Fetal bovine serum (FBS) was from Hyclone. Biochemicals were from Sigma or Life Technologies unless indicated.
Methocult m3234 methylcellulose medium
Manufactured by STEMCELL
Sourced in Canada
MethoCult M3234 is a methylcellulose-based medium designed for the in vitro culture of hematopoietic progenitor cells. It provides a semisolid matrix that supports the growth and differentiation of various cell types, including colony-forming units (CFUs) of erythroid, granulocytic, macrophage, and multipotential progenitor cells.
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Lab products found in correlation
2 protocols using methocult m3234 methylcellulose medium
1
Murine Hematopoietic Stem Cell Isolation
2
Hematopoietic Transformation Assay
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Human MOZ-TIF2, MLL-AF9, MLL-AF10, or HOXA9 genes were introduced into cells using a retrovirus system and cultured for 5 days. Green fluorescent protein (GFP) marker-positive cells were sorted by JSAN flow cytometry. Sorted GFP+ cells (5 × 104) were cultured in Methocult M 3234 methylcellulose medium (Stemcell Technologies; Vancouver, BC, Canada) containing IL-3 (20 ng/mL), SCF (50 ng/mL), GM-CSF (10 ng/mL), and 100 U/mL penicillin/streptomycin. After 3 to 5 days, colonies and total cells were counted, and 3 × 104 or 1 × 105 cells were cultured in methylcellulose medium 3 or 5 times. Cultures were maintained at 37°C in a 5% CO2 humidified atmosphere.
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