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Can professional phagocytes be easilly transfected? - (Jul/15/2007 )

What is your favarate transfection reagents when dealling with monocytes and macrophage?

-cell cracker-

QUOTE (cell cracker @ Jul 15 2007, 09:57 AM)
What is your favarate transfection reagents when dealling with monocytes and macrophage?


Primary macrophages are very hard to transfect with conventional lipid-based methods, so I would use viral vectors. Adenovirus transfects 30-50% and lentivirus will transfect close to 90%.

-mateo-

QUOTE (mateo @ Jul 16 2007, 07:09 PM)
QUOTE (cell cracker @ Jul 15 2007, 09:57 AM)
What is your favarate transfection reagents when dealling with monocytes and macrophage?


Primary macrophages are very hard to transfect with conventional lipid-based methods, so I would use viral vectors. Adenovirus transfects 30-50% and lentivirus will transfect close to 90%.



I mean macrophage-like cell-line: u937,raw,etc.Does virus-based transfection need special p3 lab? I have no experience on that.

-cell cracker-

QUOTE (cell cracker @ Jul 16 2007, 10:57 PM)
QUOTE (mateo @ Jul 16 2007, 07:09 PM)
QUOTE (cell cracker @ Jul 15 2007, 09:57 AM)
What is your favarate transfection reagents when dealling with monocytes and macrophage?


Primary macrophages are very hard to transfect with conventional lipid-based methods, so I would use viral vectors. Adenovirus transfects 30-50% and lentivirus will transfect close to 90%.



I mean macrophage-like cell-line: u937,raw,etc.Does virus-based transfection need special p3 lab? I have no experience on that.


I've transfected RAWs but only with reporter constructs. The transfection efficiency was around 10% using Fugene6. Although the lentiviral transfection is much better and BSL2, you can always make stables the conventional way depending on what your application is.

-mateo-