ShRNA (bitter taste receptor sub type konkdown in adult cardiomyocytes cells, its not reached successfully transfected based on fluorescence expression and mRNA quantification by QPCR?
Give some suggestion and feedbacks
Dear selvaraj,
If you can provide more detailed info. about whole transfection condition, definitely will get better answer. Still, overall you can re-work out these factors such as transfection reagent efficiency, preference to serum-free condition, cell viability & health.
Dear Saurabh
Thank you. Selvaraj asked this question for me.
Adult rat cardiomyocytes were maintained in vitro with M199 containing 2 mM L-glutamine, 5 mM creatine, 2 mM cartinine, 5 mM taurine and 0.8 mM EGTA.
1. They were plated in laminin-coated confocal dishes and initial attachment was performed with M199 + 10% FBS+2% of penicillin - steptomycin for 1h.
2. Remove the medium , add M199 + 2% of penicillin - steptomycin and kept in incubator for 6h.
3. Cells were incubated with lentivirus which was contained in the M199 for 12h.
4. After that, lentivirus was removed and cells were maintained with M199 + 2% of penicillin - steptomycin for another 72 h.
Cell viability was not good. After incubated with lentivirus for 72h, most cells rounded and 30-40% had rod shape.(Con-1, from control group; 70-2, from transfected group)
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