How do I address the incredible difficulty of transfecting hematopoetic cells, specifically immune system cells (e.g., macrophages such as J774A.1 and THP-1)?
This question is part of the following Ask The Expert session:
Transfection Optimization for Improved Efficiency and Performance
Company: Life Technologies
Do you really need to use this cell type?
These notoriously difficult cells require a certain amount preparation, because a variety of delivery techniques might need to be tested for success. If you are trying to over express a gene with DNA delivery, the newly developed Lipofectamine® 3000 can be tried first for the simplicity of using a lipid based delivery method. However, if this method doesn’t yield any results, the Neon™ Electroporation system with DNA is recommended. You could also try to do electroporation with an mRNA transcript of your gene of interest, which only needs to be delivered to the cytoplasm for protein translation. If all these previous methods fail, a virus can be used for gene expression.
If gene silencing with siRNA is desired, the lipid based Lipofectamine® RNAiMax can be tested; however, the Neon™ Electroporation system is the recommended method. If this proves unsuccessful, an shRNA vector can be delivered.