This problem drives me crazy for over a year now. I have tried a couple of non-viral transfection agents including PEI, fusogenic liposomes, and electroporation but none of these methods gives me a satisfying efficiency. I was wondering if there is something special about cardiomyocytes that makes them less prone to transfection (e.g. different membrane potential, glycocalyx or plasma membrane composition).
Hi Benjamin,
those cells are indeed extremely difficult to transfect for a still unclear reason. one Option for you would be to try Magnetofection.
http://www.ozbiosciences.com/content/13-magnetofection
you can look at the citation database
http://www.ozbiosciences.com/module/citationfinder/default
It has been used successfully for these cells with the reagent called NeuroMag and we just receive very positive results from another customer
here is the publication: http://www.ncbi.nlm.nih.gov/pubmed/23936108
Best regards,
Olivier
You are right, and based on our experience, it is relatively easier to transfect embryonic stem cell-derived neurons with non-viral transfection agents than hESC-derived cardiomyocytes. Generally, non-viral transfection agents have low transfection efficiency on differentiated and non proliferative cells than specific types of virus.
Dear Benjamin,
indeed transfection of cardiomyocytes is a very hard challenge.
I would advise to try Viromer RED or YELLOW for plasmid/shRNA transfection and Viromer BLACK for siRNA/miRNA transfection (please find data attached).
These reagents were developed from Lipocalyx (www.lipocalyx.de).
Best,
Olivia
Benjamin, We are wondering about this too, as we have made genetically-encoded fluorescent sensors for DAG, cAMP and voltage that we want to try in cardiomyocytes (Cor.4u)
Since all of the sensors are in BacMam, we'll try a range of amounts to optimize expression. BacMam is a modified virus so it doesn't replicate (BSL-1) and it doesn't integrate into the genome.
Anne Marie
Article New DAG and cAMP Sensors Optimized for Live-Cell Assays in A...
- Badges
- Science method