Hi Everyone my PI was looking into isolating exosomal miRNA from MC3T3 cell lines. We learnt that before we can even begin this project we would need exosomal free media which cost around $1400. I was wondering if there was a cheaper alternative, perhaps using an ultracentrifuge or something.
Unfortunately, ultracentrifugation at 100k for 2-18 hours only removes 19-33% of RNA content in FBS. Article Fetal Bovine Serum RNA Interferes with the Cell Culture deri...
You should include control such as complete growth media that isn't conditioned with MC3T3 cells in your analysis.
Yes, there are simple and cheaper ways to remove exosomes from small volumes of serum. Please see link below. However, a complete removal of exosomes from serum is not possible. Therefore, a control that Nghiem suggested would be needed.
https://systembio.com/products/exosome-research/exosome-isolation/
Hi Omar Khan ,
1. Practically it is not possible to completely remove the exosomes from any media or culture.
2. Exosomes are the vesicles of 10-100 nm size.
3. You can minimize the count by using milipore filter of minimumsize as er your choice.
4. Further removal can be done bu ultracentrifugation at very high speed (more than 100000 rpm) for 2-10 hours.
5. Moreover, you to standardized all these exercises in your lab condition as per your requirement.
Thank you,
Good luck