For simulated ischemia, growth or preconditioning media was replaced with saline to mimic nutrient/energy deprivation and placed in an airtight chamber flushed with nitrogen gas at 37°C for 4hrs. Saline supernatant was removed after simulated ischemia for LDH measurement. Reperfusion was simulated by adding back fresh complete DMEM-12320 (Invitrogen) containing 0.5 mg/mL MTT under normoxia at 37°C for an additional 3–4hrs.
Dmem 12320
Manufactured by Thermo Fisher Scientific
DMEM-12320 is a cell culture medium manufactured by Thermo Fisher Scientific. It is a formulation of Dulbecco's Modified Eagle's Medium, which is commonly used to support the growth and maintenance of a variety of cell types in vitro.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using dmem 12320
1
Simulated Ischemia and Reperfusion Assay
2
Isolation and Culturing of MEFs
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Mouse embryonic fibroblasts (MEFs) were isolated by female wt and Crh
−/− mice, when embryos were
13.5–14.5 days old as previously described64 (link).
Isolated fibroblasts were cultured in Dulbecco’s modified
Eagle’s medium (DMEM) with 10% fetal bovine serum (FBS), 1%
penicillin/streptomycin and 1% L-glutamine. For the aminoacid deprivation
experiments, cells were incubated for 4 h with a special formulation
of Invitrogen DMEM-12320. RAW264.7 cells were maintained in
Dulbecco’s modified Eagle’s medium containing 10% FBS.
For LPS induced autophagy experiments, RAW264.7 cells were treated with LPS
(1 μg/ml, L2630, Sigma Aldrich), Crh
(10−7M, C3042, Sigma Aldrich) or both.
−/− mice, when embryos were
13.5–14.5 days old as previously described64 (link).
Isolated fibroblasts were cultured in Dulbecco’s modified
Eagle’s medium (DMEM) with 10% fetal bovine serum (FBS), 1%
penicillin/streptomycin and 1% L-glutamine. For the aminoacid deprivation
experiments, cells were incubated for 4 h with a special formulation
of Invitrogen DMEM-12320. RAW264.7 cells were maintained in
Dulbecco’s modified Eagle’s medium containing 10% FBS.
For LPS induced autophagy experiments, RAW264.7 cells were treated with LPS
(1 μg/ml, L2630, Sigma Aldrich), Crh
(10−7M, C3042, Sigma Aldrich) or both.
3
Isolation and Culture of Dermal Fibroblasts and Hepatocytes
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