Suspension:
Article Title: Attenuating amyloid-beta pathology in mice with in situ programmed astrocytes
Article Snippet: .. For astrocyte isolation, 100 μl of 0.5 mg/ml sheep anti-ITGB5 (R&D Systems, AF3824) was added into 5-10 ml of cell suspension and incubated with the cells for 30-40 min at 37°C, and cell suspensions were allowed to interact with the immunopanning dish (coated with 60 μl of secondary antibodies in 50 mM Tris-HCl pH 9.5 overnight at 4°C) for 20 min at room temperature, unbound cells and debris were removed by washing the dish with dPBS ten consecutive times, freshly isolated astrocytes were cultured in DMEM/Neurobasal (1:1) containing 100 units/mL penicillin, 100 μg/mL streptomycin (ThermoFisher Scientific,15140122), 1 mM sodium pyruvate (ThermoFisher Scientific, 11360070), 292 μg/ml L-glutamine (ThermoFisher Scientific, 25030081), 1 μg/mL transferrin (Sigma, T8158), 0.16 μg/mL putrescine (Sigma, P5780), 1 nM progesterone (Merck, P0130), 0.4 ng/ml sodium selenite (ThermoFisher Scientific, 11360070), 5 μg/ml N-Acetyl-L-cysteine (Sigma, A8199) and 5ng/ml HBEGF (MedChemExpress, #HY-P7400). .. For microglia isolation, cell suspensions were applied directly to positive-selection immunopanning dishes coated with anti-CD11b monoclonal antibodies, freshly isolated microglia were cultured in DMEM/F12 containing 100 units/mL penicillin, 100 μg/mL streptomycin, 2 mM L-glutamine, 5 μg/ml N-acetyl cysteine, 5 μg/ml insulin (Merck, I9278), 100 μg/mL transferrin, and 100 ng/mL sodium selenite.
Incubation:
Article Title: Attenuating amyloid-beta pathology in mice with in situ programmed astrocytes
Article Snippet: .. For astrocyte isolation, 100 μl of 0.5 mg/ml sheep anti-ITGB5 (R&D Systems, AF3824) was added into 5-10 ml of cell suspension and incubated with the cells for 30-40 min at 37°C, and cell suspensions were allowed to interact with the immunopanning dish (coated with 60 μl of secondary antibodies in 50 mM Tris-HCl pH 9.5 overnight at 4°C) for 20 min at room temperature, unbound cells and debris were removed by washing the dish with dPBS ten consecutive times, freshly isolated astrocytes were cultured in DMEM/Neurobasal (1:1) containing 100 units/mL penicillin, 100 μg/mL streptomycin (ThermoFisher Scientific,15140122), 1 mM sodium pyruvate (ThermoFisher Scientific, 11360070), 292 μg/ml L-glutamine (ThermoFisher Scientific, 25030081), 1 μg/mL transferrin (Sigma, T8158), 0.16 μg/mL putrescine (Sigma, P5780), 1 nM progesterone (Merck, P0130), 0.4 ng/ml sodium selenite (ThermoFisher Scientific, 11360070), 5 μg/ml N-Acetyl-L-cysteine (Sigma, A8199) and 5ng/ml HBEGF (MedChemExpress, #HY-P7400). .. For microglia isolation, cell suspensions were applied directly to positive-selection immunopanning dishes coated with anti-CD11b monoclonal antibodies, freshly isolated microglia were cultured in DMEM/F12 containing 100 units/mL penicillin, 100 μg/mL streptomycin, 2 mM L-glutamine, 5 μg/ml N-acetyl cysteine, 5 μg/ml insulin (Merck, I9278), 100 μg/mL transferrin, and 100 ng/mL sodium selenite.
Isolation:Article Title: Attenuating amyloid-beta pathology in mice with in situ programmed astrocytes
Article Snippet: .. For astrocyte isolation, 100 μl of 0.5 mg/ml sheep anti-ITGB5 (R&D Systems, AF3824) was added into 5-10 ml of cell suspension and incubated with the cells for 30-40 min at 37°C, and cell suspensions were allowed to interact with the immunopanning dish (coated with 60 μl of secondary antibodies in 50 mM Tris-HCl pH 9.5 overnight at 4°C) for 20 min at room temperature, unbound cells and debris were removed by washing the dish with dPBS ten consecutive times, freshly isolated astrocytes were cultured in DMEM/Neurobasal (1:1) containing 100 units/mL penicillin, 100 μg/mL streptomycin (ThermoFisher Scientific,15140122), 1 mM sodium pyruvate (ThermoFisher Scientific, 11360070), 292 μg/ml L-glutamine (ThermoFisher Scientific, 25030081), 1 μg/mL transferrin (Sigma, T8158), 0.16 μg/mL putrescine (Sigma, P5780), 1 nM progesterone (Merck, P0130), 0.4 ng/ml sodium selenite (ThermoFisher Scientific, 11360070), 5 μg/ml N-Acetyl-L-cysteine (Sigma, A8199) and 5ng/ml HBEGF (MedChemExpress, #HY-P7400). .. For microglia isolation, cell suspensions were applied directly to positive-selection immunopanning dishes coated with anti-CD11b monoclonal antibodies, freshly isolated microglia were cultured in DMEM/F12 containing 100 units/mL penicillin, 100 μg/mL streptomycin, 2 mM L-glutamine, 5 μg/ml N-acetyl cysteine, 5 μg/ml insulin (Merck, I9278), 100 μg/mL transferrin, and 100 ng/mL sodium selenite.
Cell Culture:Article Title: Attenuating amyloid-beta pathology in mice with in situ programmed astrocytes
Article Snippet: .. For astrocyte isolation, 100 μl of 0.5 mg/ml sheep anti-ITGB5 (R&D Systems, AF3824) was added into 5-10 ml of cell suspension and incubated with the cells for 30-40 min at 37°C, and cell suspensions were allowed to interact with the immunopanning dish (coated with 60 μl of secondary antibodies in 50 mM Tris-HCl pH 9.5 overnight at 4°C) for 20 min at room temperature, unbound cells and debris were removed by washing the dish with dPBS ten consecutive times, freshly isolated astrocytes were cultured in DMEM/Neurobasal (1:1) containing 100 units/mL penicillin, 100 μg/mL streptomycin (ThermoFisher Scientific,15140122), 1 mM sodium pyruvate (ThermoFisher Scientific, 11360070), 292 μg/ml L-glutamine (ThermoFisher Scientific, 25030081), 1 μg/mL transferrin (Sigma, T8158), 0.16 μg/mL putrescine (Sigma, P5780), 1 nM progesterone (Merck, P0130), 0.4 ng/ml sodium selenite (ThermoFisher Scientific, 11360070), 5 μg/ml N-Acetyl-L-cysteine (Sigma, A8199) and 5ng/ml HBEGF (MedChemExpress, #HY-P7400). .. For microglia isolation, cell suspensions were applied directly to positive-selection immunopanning dishes coated with anti-CD11b monoclonal antibodies, freshly isolated microglia were cultured in DMEM/F12 containing 100 units/mL penicillin, 100 μg/mL streptomycin, 2 mM L-glutamine, 5 μg/ml N-acetyl cysteine, 5 μg/ml insulin (Merck, I9278), 100 μg/mL transferrin, and 100 ng/mL sodium selenite.
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