Bone marrow derived MSCs were isolated from the femoral shaft of 6 week old rats (Fischer Male) and expanded in high-glucose Dulbecco's modified Eagle's medium GlutaMAX (hgDMEM) supplemented with 10% v/v fetal bovine serum (FBS), 100 U/ml penicillin, 100 μg/ml streptomycin (all Gibco Biosciences, Dublin, Ireland) and 2.5 μg/ml amphotericin B (Sigma-Aldrich, Dublin, Ireland) at 5% pO2. Following colony formation, MSCs were trypsinized, counted, seeded at density of 5000 cells cm 2 in 500 cm 2 triple flasks (Thermo Fisher Scientific), supplemented with hgDMEM, 10% v/v FBS, 100 U/ml penicillin, 100 μg/ml streptomycin, 2.5 μg/ml amphotericin B, and 5 ng/ml human fibroblastic growth factor-2 (FGF-2; Prospec-Tany TechnoGene Ltd., Israel) and expanded to passage 2 at 5% pO2.
Amphotericin b
Manufactured by ProSpec
Sourced in Israel
Amphotericin B is a polyene antifungal medication. It is a naturally occurring compound produced by the bacterium Streptomyces nodosus. Amphotericin B is used as a laboratory reagent in cell culture and microbiology applications.
Automatically generated - may contain errors
2 protocols using amphotericin b
1
Rat Bone Marrow Mesenchymal Stem Cell Isolation
2
Isolation and Expansion of Porcine Mesenchymal Stem Cells
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Bone marrow–derived MSCs were isolated from the femoral shaft of 4-month-old pigs and expanded according to a modified method for human MSCs26 (link) in high-glucose Dulbecco’s modified Eagle’s medium (hgDMEM) GlutaMAX supplemented with 10% v/v fetal bovine serum (FBS), 100 U mL−1 penicillin/100 μg mL−1 streptomycin (all Gibco Biosciences, Dublin, Ireland) and 2.5 μg mL−1 amphotericin B (Sigma-Aldrich, Dublin, Ireland) at 20% pO2. Tri-potentiality was confirmed prior to use. Following colony formation, MSCs were trypsinised, counted, seeded at density of 5000 cells cm2 in 500 cm2 (link) triple flasks (Thermo Fisher Scientific), supplemented with hgDMEM, 10% v/v FBS, 100 U mL−1 penicillin/100 μg mL−1 streptomycin, 2.5 μg mL−1 amphotericin B and 5 ng mL−1 human fibroblastic growth factor-2 (FGF-2; ProSpec-Tany TechnoGene Ltd, Israel) and expanded to passage 2.
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