I am growing Mouse Prostate organoids in the Matrigel domes and they achieve their appropriate size after 7 days.
I tried to transfect full grown organoids at 7 day using si3D-FectIN , but it didn’t work as si3D-FectIN could not penetrate the polymerised matrigel. I am using BLOCK-iT™ Alexa Fluor® Red Fluorescent as a Control (50nM).
Then i seeded the cells in the matrigel along with the si3D-FectIN transfection complex same day.The organoids started to grow, although less in number and size.
However i could not see any fluorescence in the bigger organoids at 3,4,5,6,7 th day. However i could see fluorescence in very small organoids.
How to increase transfection efficiency especially in case of bigger organoids?
Hi Sajad,
I am not an expert in this but did you already check the below article about how the presence of serum in the transfection mix helps transfection efficiency in 3D cultures?
https://www.nature.com/articles/s41598-018-26253-3
Thanks
Satish
Hi Satish,
Yeah i got to know this few days before. I have attempted to transfect organoids in presence of serum. It is working to some extent. i am optimising the conditions to improve transfection efficiency using this protocol.
Thanks
Sajad
Satish Bodakuntla