We isolated exosomes from culture supernatants using the ultracentrifugation method. Could anyone suggest how to transfect these exosomes and purify them for consecutive administration in vivo?
Dear Steffi,
I guess one of the best method will be to use the magnetofection technology that creates magnetic complexes only on the basis of electrostatic interactions and allow concentration onto the cell surface under a specific magnetic field.
This transfection method was demonstrated to be highly efficient in numerous primary cells and hard-to-transfect cells, for any vector (DNA, siRNA, mRNA, lentivirus, adenovirus...) and can be also applied in vivo. Actually it is the only method that allow targeting transfection in vivo.
It couls be really interesting to try this and as we are located in Marseille it could be really easy to find a solution.
You can contact me at [email protected] if you need any supplemental information.
best regards,
Cedric
Dear Bosch,
I am transfecting cells with Lentivirus that contain a self-made plasmid which targets miRNAs and antimiRNAs to the Exosomes. After that, I am isolating the exosomes from the cells by ultracentrifuge under sterilized conditions for in vivo administration.
If do you have any question please feel free to ask,
Best regards,
we loaded exosomes with mirVana miRNA mimics and inhibitors, and Silencer Select siRNA- using Neon electroporation (tried many parameters). but efficiency was extremely low. despite several papers are published on exosome electroporation and other loading techniques- none of them work well unfortunately. not yet
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