Human CD34+ cells were cultured in Stemspan serum-free medium II (SFEM II, StemCell Technologies), supplemented with granulocyte-macrophage colony-stimulating factor (GM-CSF) 200pg/mL, leukemia inhibitory factor (LIF) 50pg/mL, granulocyte colony-stimulating factor (G-CSF) 1ng/mL, stem cell factor (SCF) 200pg/mL, macrophage-inflammatory protein-1α (MIP-1α) 200pg/mL, and interleukin-6 (IL-6) 1ng/mL.23 (link)–25 (link) Mouse BM Lineage−Sca-1+c-Kit+ (LSK) cells were cultured in SFEM II supplemented with 10ng/ml SCF and 10ng/ml TPO.25 (link) All cytokines were obtained from Pepro Tech US (Rocky Hill, NJ, USA).
Protocol detail
Culture of Human and Mouse Hematopoietic Stem Cells
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Cells
Cultured medium
Cytokines
G csf
Granulocyte macrophage colony stimulating factor
Human
Interleukin 23
Leukemia inhibitory factor
Macrophage inflammatory protein 1α
Mouse
Sca 1
Serum
Stem cell factor
Stemcell
Corresponding Organization :
Other organizations : Zhejiang University, City of Hope
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Variable analysis
independent variables
- Culture medium (Stemspan serum-free medium II (SFEM II, StemCell Technologies))
- Cytokine supplementation: granulocyte-macrophage colony-stimulating factor (GM-CSF) 200pg/mL, leukemia inhibitory factor (LIF) 50pg/mL, granulocyte colony-stimulating factor (G-CSF) 1ng/mL, stem cell factor (SCF) 200pg/mL, macrophage-inflammatory protein-1α (MIP-1α) 200pg/mL, and interleukin-6 (IL-6) 1ng/mL
- Cell types: Human CD34+ cells, Mouse BM Lineage-Sca-1+c-Kit+ (LSK) cells
- Measured outcomes not explicitly mentioned
- Cytokine concentrations
- Culture medium (SFEM II)
- Positive control: Not specified
- Negative control: Not specified
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