Colony formation assays were performed as described previously 8 . HL60, MOLM13 and MV411 cells were treated with varying doses of S-250 for 4 hours. After 4 hours, cells were washed to remove the drug and plated in media containing 20% serum and 0.3% soft agar (Noble agar, Sigma). Colonies were allowed to form for 10 days and were counted under a light microscope. For normal bone marrow samples, cells were treated with 500nM S-250 for 4h, washed and plated on methylcellulose (Methocult Classic, Stem cell technologies) containing 0.25mM hemin (Sigma). For AML patient samples, cells were likewise treated for 4h with 500nM S-250 and seeded on methylcellulose supplemented with 20% serum and 20ng/ml G-CSF (GoldBio). Colonies were counted after incubating for 10 days.
G csf
Manufactured by GoldBio
G-CSF is a recombinant human granulocyte colony-stimulating factor. It is a protein that stimulates the production and function of neutrophils, a type of white blood cell. G-CSF plays a crucial role in the regulation of the immune system and hematopoiesis.
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2 protocols using g csf
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Colony Formation Assays for AML Cells
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Colony Formation Assays for AML Cells
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Colony formation assays were performed as described previously 8 . HL60, MOLM13 and MV411 cells were treated with varying doses of S-250 for 4 hours. After 4 hours, cells were washed to remove the drug and plated in media containing 20% serum and 0.3% soft agar (Noble agar, Sigma). Colonies were allowed to form for 10 days and were counted under a light microscope. For normal bone marrow samples, cells were treated with 500nM S-250 for 4h, washed and plated on methylcellulose (Methocult Classic, Stem cell technologies) containing 0.25mM hemin (Sigma). For AML patient samples, cells were likewise treated for 4h with 500nM S-250 and seeded on methylcellulose supplemented with 20% serum and 20ng/ml G-CSF (GoldBio). Colonies were counted after incubating for 10 days.
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