Dear All,
Does anyone has a positive experience with MM1.S or/and U266 multiple myeloma cells transfection. I need to transfect these cells and not to electroporate and now looking for a reagent that can give more then 30% cells transfected. I appreciate any suggestion.
Elena
Our lab was able to ton transfect 8226, u266 and mm1s. They are extremely difficult to do, If you can use retrovirus, that is your best bet.
Otherwise, I would recommend including a fluorescent marker to enable FACS sorting a couple days following exposure. I would expect to perform multiple ficoll purifications if doing drug selection. If transient, Amaxa company has protocols already worked out with their electroporator. $$$$$ but pretty stunning results.
We had mild success with cationic lipids. Keep the cells dense (but don't let over grow). DNA on the low end, transfection reagent in the mid range. If I get time I'll try to get you a specific protocol. If can't do virus try the amaxa systems
right they are now called lanza, you'll see your be cell protocols on their website. http://www.lonza.com/products-services/bio-research/transfection/nucleofector-technology.aspx
Hello, do you know the Viromer technology? Our reagents are polymer-based chemicals mimicking a viral process of endosome escape. They can give pretty good results for hard-to-transfect cells. Unfortunately, there is no feedbacks until now for your specific cells, but we would be more than happy to get yours. In general, it works well with cancer cell lines.
Please, tell me, depending on your needs (siRNA, pDNA, mRNA transfection), I can organize free test samples for you. We have a distributor in Israel.
Bests,
Sandra
(below, links for the technology and our cell database)
https://viromer-transfection.com/technology
https://viromer-transfection.com/data-by-cell-type/cell-transfection-a-z
Hi Elena - I transduce these cells regularly using lentivirus and normally get 90-95% kd the following methods...
1 - 50k cells per well, in 100ul of RPMI media (no p/s, 10%BSA)
2 - I normally use an MOI of around 30 for MM cell lines, add polybrene to aid transduction
3 - After 24 hours, pool all wells of the same kd and add more media to help dilute the virus (for 100ul add at least another 200ul for example)
4 - test for kd at 96 hours
The kd is normally good for 2-3 weeks.
Best of luck
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