I am wondering if there is an effective transfection protocol, that increases the efficiency in this suspension cells. Any tips are welcome.
We used Neon transfection system (Invitrogen). It works OK but not that great.
Good luck
Ram
We have used multiple methods to introduce plasmids containting target cDNA or siRNA or shRNA into multiple myeloma cell lines. These methods include Amaxa Nucleofector, lipofectamine, and electroporation, and retrovirus vector expression system and Lentivirus vector expression system. The highest transfection (infection) efficiency among these methods is lentivirus vector expressing system, the gift from lab of Prof. Didier Trono (Université de Genève, Switzerland). Besides the high efficiency, cDNA in lentivirus expressing vector integrates with chromosome of the cells and thus prevents leaking of ectopic expressing genes in the vector from the expressed cells after culture of myeloma cells in vitro for long time.
I agree with Ya-Wei, electroporation or lentiviral are best. We have also had some success with streptolysin-O
Hi Vianihuini,
I am also facing a similar problem in transfecting myeloma cell lines. I have tried all lipid based reagents and non of them have worked. I am now trying Amaxa nucleofection and trying to optimize the best program. I have tried those settings recommended by Amaxa, but all of my cells were killed. So I am now trying different settings. May I know which myeloma cell lines are you transfecting? I am using RPMi 8226 and U266.
I will update you as I have some positive results.
Cheers,
Definitely lentivirus. But you will need containment level 2 labs to do this.
If you must use transfection, see this paper:
http://www.ncbi.nlm.nih.gov/pubmed/24901949
Good luck.
Hi, Amaxa nucleofection works well with these cells. I am using Amaxa program T-001 for both the cell lines and getting quite a good knock down.
Good luck,
For me, the Viromer Red/Yellow did not work. They gave a similar transfection efficiency to lipofectamine 2000 (0.5-1% max). However, the best results for me was Amaxa. For U266 I used Amaxa program X-005 and MM1S I used O-023. T-001 led to a lot of cell death in RPMI8226 for me. Maybe a nucleofection buffer issue.
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