StemSpan™ SFEM

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	semisolid colony-forming serum-free medium (CF-SFM) containing 40% ES-Cult™ M3120 methylcellulose (2.5% solution in IMDM; Stem Cell Technologies, Vancouver, BC, Canada), 25% StemSpan™ serum-free expansion medium (SFEM; Stem Cell Technologies), 25% human endothelial serum-free medium (ESFM; Invitrogen), 10% BIT 9500 supplement (Stem Cell Technologies), GlutaMAX™ (1/100 dilution; Invitrogen), Ex-Cyte® supplement (1/1000 dilution; Millipore, Billerica, MA), 100 μM MTG, 50 μg/ml ascorbic acid and 20 ng/ml basic fibroblast growth factor (FGF2)	human PDGF-BB (10 ng/ml), VEGF (20 ng/ml), stem cell factor (SCF; 50 ng/ml), epidermal growth factor (EGF; 20 ng/ml), insulin-like growth factor 1 (IGF1; 50 ng/ml), hepatocyte growth factor (HGF; 20 ng/ml), activin A (1 ng/ml), transforming growth factor β1 (TGF-β1; 1 ng/ml), bone-morphogenic protein-4 (BMP-4; 5 ng/ml), apelin-12 (100 ng/ml; Phoenix Pharmaceuticals Inc), or anti-APLNR monoclonal antibody (5 μg/ml; R&D Systems) were added to cultures

Protocol tips
semisolid colony-forming serum-free medium (CF-SFM) containing 40% ES-Cult™ M3120 methylcellulose (2.5% solution in IMDM; Stem Cell Technologies, Vancouver, BC, Canada), 25% StemSpan™ serum-free expansion medium (SFEM; Stem Cell Technologies), 25% human endothelial serum-free medium (ESFM; Invitrogen), 10% BIT 9500 supplement (Stem Cell Technologies), GlutaMAX™ (1/100 dilution; Invitrogen), Ex-Cyte® supplement (1/1000 dilution; Millipore, Billerica, MA), 100 μM MTG, 50 μg/ml ascorbic acid and 20 ng/ml basic fibroblast growth factor (FGF2)
Downstream tips
human PDGF-BB (10 ng/ml), VEGF (20 ng/ml), stem cell factor (SCF; 50 ng/ml), epidermal growth factor (EGF; 20 ng/ml), insulin-like growth factor 1 (IGF1; 50 ng/ml), hepatocyte growth factor (HGF; 20 ng/ml), activin A (1 ng/ml), transforming growth factor β1 (TGF-β1; 1 ng/ml), bone-morphogenic protein-4 (BMP-4; 5 ng/ml), apelin-12 (100 ng/ml; Phoenix Pharmaceuticals Inc), or anti-APLNR monoclonal antibody (5 μg/ml; R&D Systems) were added to cultures

Publication protocol

semisolid colony-forming serum-free medium (CF-SFM) containing 40% ES-Cult™ M3120 methylcellulose (2.5% solution in IMDM; Stem Cell Technologies, Vancouver, BC, Canada), 25% StemSpan™ serum-free expansion medium (SFEM; Stem Cell Technologies), 25% human endothelial serum-free medium (ESFM; Invitrogen), 10% BIT 9500 supplement (Stem Cell Technologies), GlutaMAX™ (1/100 dilution; Invitrogen), Ex-Cyte® supplement (1/1000 dilution; Millipore, Billerica, MA), 100 μM MTG, 50 μg/ml ascorbic acid and 20 ng/ml basic fibroblast growth factor (FGF2)

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Papers

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Manufacturer protocol

Download the product protocol from STEMCELL technologies for StemSpan™ SFEM below.

Download manufacturer protocol

Videos

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