Isolation and Culture of Adult Mouse Cardiomyocytes

Cardiomyocytes CMs were isolated from adult mouse hearts (8–10 weeks of age) using a simplified Langendorff-free method^{78 (link)} and cultured in M199 (Sigma, M4530) supplemented with 0.1% BSA (supplier, number), 1X ITS Liquid Media Supplement (Sigma, I3146), 10 mmol/l BDM (Sigma, B0753), 1X Chemically Defined Lipid Concentrate (Fisher, 11548846) and penicillin–streptomycin (Thermo Fisher, 15140-122). When needed, adenovirus was administered to CMs in culture media immediately after removal of plating media. Medium was then replaced after 24 h with culture media together with 100 nM (Z)-4-OHT (Sigma, H7904) in ethanol and collected after 4 h for RNA extraction.

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Bywater M.J., Burkhart D.L., Straube J., Sabò A., Pendino V., Hudson J.E., Quaife-Ryan G.A., Porrello E.R., Rae J., Parton R.G., Kress T.R., Amati B., Littlewood T.D., Evan G.I, & Wilson C.H. (2020). Reactivation of Myc transcription in the mouse heart unlocks its proliferative capacity. Nature Communications, 11, 1827.

Publication 2020

BSA ITS Liquid Media Supplement penicillin–streptomycin Z)-4-OHT

4 oht Adenovirus Adult Cardiomyocytes Culture media Ethanol Hearts Lipid Mouse Penicillin Streptomycin Supplement Z 100

Corresponding Organization : University of Cambridge

Other organizations : QIMR Berghofer Medical Research Institute, European Institute of Oncology, Italian Institute of Technology, Center for Genomic Science, Royal Children's Hospital, Murdoch Children's Research Institute, University of Queensland

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Variable analysis

independent variables

Administration of adenovirus to cardiomyocytes (CMs) in culture media

dependent variables

RNA extraction after 4 hours

control variables

Culture of cardiomyocytes (CMs) in M199 medium supplemented with 0.1% BSA, 1X ITS Liquid Media Supplement, 10 mmol/l BDM, 1X Chemically Defined Lipid Concentrate, and penicillin-streptomycin
Replacement of culture media with 100 nM (Z)-4-OHT in ethanol after 24 hours

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

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