Hi all, I am appreciated if someone could suggest the duration for oxygen glucose deprivation (OGD) to mimic the ischemia/reperfusion injury for H9c2 cell line. Thanks,
This is something only you can decide because you must have in mind the level of deprivation you intend to mimic.
The method is to make kinetic ischemia with different times. Depending on what you seek to show, you can later adapt the time to your model. For example, if you want to get 30% of apoptotic cells, you can perform kinetics 15min, 1h, 2h, 4h, 8h. Then you have to analyze your cells (viability, marking microscopy, cytometry, cleaved caspase-3 in western blot, etc.).
Good luck !
Hi Nghiem Le,
I agree with Tausif and Helene's comments.
Initially, it may be beneficial to undertake some optimisation studies investigating the effect of time on rendering the cells temporarily ischaemic.
My own experience in this area has involved undertaking in-vivo studies, recruiting orthopaedic patients undergoing upper and lower limb surgery, where their limbs have been rendered temporarily ischaemic for various times, ranging from ~15-95 minutes.
Refs:
Hughes SF, Hendricks BD, Edwards DR, & Middleton JF, (2010). Tourniquet-applied upper limb orthopaedic surgery results in increased inflammation and changes to leukocyte, coagulation & endothelial markers. PLOS ONE 5(7): e11846. doi:10.1371/journal.pone.0011846.
Stephen F. Hughes, Beverly D. Hendricks, David R. Edwards, Kirsty M. Maclean, Salah S. Bastawrous, & Jim F. Middleton (2010). Total hip and knee replacement surgery results in changes in leukocyte and endothelial markers. Journal of Inflammation [Lond.] 7(2) doi:10.1186/1476-9255-7-2.
Good luck.
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