Do you have a protocol for BacMam/AAV/Lenti gene delivery in iCell Cardiomyocytes?

We have tested the utility of the BacMam system to endogenously deliver genes such as GFP into our iCell® Cardiomyocytes; we have not tested infection using AAV or lentivirus, but our customers have successful experience with both of these methods.

We currently do not have an optimized Application Protocol for BacMam/AAV/lentivirus transduction, but we can provide general guidelines to help you get started (e.g., always run functional assays two days after transduction) and recommendations to optimize the procedure for your gene of interest. For additional information and troubleshooting steps, we recommend that you consult the vendor.

General protocol recommendations:

96-well option

  1. Plate cells in 96-well vessels and culture according to our User’s Guide.
  2. On day 8 post-thaw, transduce the cells with BacMam. Try a 10% virus load by volume (v/v) with and without Enhancer Solution (0.2X final concentration, diluted from a 1 mM (1000X) stock solution).
  3. On day 9, remove the BacMam transduction solution.
  4. On day 10 (48 hours post-transduction), run functional assays.

6-well option

  1. Plate cells in 6-well vessels and culture according to our User’s Guide.
  2. On day 7 post-thaw, transduce the entire well "in batch".
  3. On day 8, dissociate the cells and re-plate into a 96-well format.
  4. On day 10, run functional assays.

References:

  1. iCell Cardiomyocytes User’s Guide
  2. Poster Presentation: Dempsey et al (SPS 2014)_Drug Discovery and Toxicity Screening in Stem Cell Models Using Optogenetics
  3. Traister et al 2014 Integrin-Linked Kinase Mediates Force Transduction in Cardiomyocytes by Modulating SERCA2a/PLN Function
  4. Liu et al 2015 EAG Domains Regulate LQT Mutant hERG Channels in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes
  5. Webinar Presentation (Promega and Viafect): Efficiently Build Relevant In Vitro Models Using Human Stem Cell-Derived Tissue Cells, High Performance Transfection and Novel Multiplexed Reporter Techniques