A cell culture model that recapitulates CS-WR was used as previously described.17 (link) Briefly, a confluent monolayer of ECs underwent a 6-hour period of cold ischemia in University of Wisconsin (UW) solution (Bridge to Life, SC) at 4°C in a sealed hypoxic chamber preflushed with pure nitrogen (Airgas, SC). After cold ischemia, UW was removed and replaced with warm culture media to stimulate reperfusion injury. For mPTP modulation, 1μM NIM811 was added to the reperfusion media.
Protocol detail
Recapitulating Cold Ischemia-Reperfusion Injury
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Cell culture
Cold ischemia
Culture media
Hypoxic
Mptp
Nim811
Nitrogen
Reperfusion
Reperfusion injury
Corresponding Organization : Medical University of South Carolina
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Variable analysis
independent variables
- Duration of cold ischemia (6 hours)
- Addition of NIM811 (1 μM) to reperfusion media
- Extent of reperfusion injury
- Cell culture model that recapitulates CS-WR
- Confluent monolayer of endothelial cells (ECs)
- University of Wisconsin (UW) solution for cold ischemia
- Temperature during cold ischemia (4°C)
- Sealed hypoxic chamber with pure nitrogen for cold ischemia
- Replacement of UW solution with warm culture media to stimulate reperfusion injury
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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