Can you transfect primary cells?
Transfection of primary cells and stem cells is a problem in the laboratory routine and further in tissue engineering and gene therapy. Most methods working effectively for cell lines in culture fail to transfect primary cells. Here we describe the use of the Nucleofector technology developed by amaxa biosystems.
What are transfection reagents?
Transfection reagents are used to introduce naked or purified nucleic acids into eukaryotic cells. These products may include plasmids, reagents, kits, primer sets, and more.
What are the two chemical reagents that can be used for transfection?
Transfection reagents can be generally divided into liposomal or high-lipid based and non-liposomal based reagents. Non-liposomal reagents can be mixed protein-lipid reagents, non-liposomal lipids or non-protein non-lipid reagents like dendrimer.
What is the best transfection reagent?
These results suggest that FuGENE HD is the most preferred transfection reagent for many cell lines, followed by Arrest-In and jetPEI. These results may be useful for improving nonviral gene and cell therapy applications.
How do you transfect cells?
The physical transfection methods are the most recent methods and use diverse physical tools to deliver nucleic acids. The methods include direct micro injection, biolistic particle delivery, electroporation, and laser-based transfection .
What is transfection method?
Transfection is the process of introducing nucleic acids into eukaryotic cells by nonviral methods. Using various chemical or physical methods, this gene transfer technology enables the study of gene function and protein expression in a cellular environment.
What are the methods of cell transfection?
Physical methods include electroporation, microinjection, gene gun, impalefection, hydrostatic pressure, continuous infusion, and sonication. Chemicals include methods such as lipofection, which is a lipid-mediated DNA-transfection process utilizing liposome vectors.
Why is opti MEM used for transfection?
OPTI-MEM is the best option since transfection efficiency is high when cells are grown in O-MEM during transfection. FBS in regular media hinders transfection so using Opti-MEM is good option when using Lipofectamine.
How do cells prepare for transfection?
Serum-containing or serum-free medium appropriate for the cell type can be used for culturing the cells. To prepare cells for transfection, remove the medium in each culture well and replace with 0.5 mL fresh complete medium.