EVB-transformed B cell transfection
K.Mulcahy at sheffield.ac.uk
Tue Feb 6 12:57:16 EST 1996
I wish to transfect tumour-associated antigens into human
EBV-transformed B cell lines. However, I have come across two main
Firstly, I understand that EBV-transformed B cell lines are notoriously
difficult to transfect. Therefore, does anybody use a transfection
method which has provided them with "satisfactory" results with
EBV-transformed B cell lines?
Secondly, if transfectants are obtained, I need a procedure to clone
them. However, using normal growth medium (RPMI/10% FCS), I have had no
success with serial-dilution cloning or soft-agar cloning using normal
growth medium. From the literature, I gather that the use of human
endothelial cell supernatant (HECS) greatly increases the ability of
EBV-transformed B cell lines to grow when there is only one or two cells
per well. Does anybody know if HECS can obtained commercially or do I
have to prepare it in-house?
Any advice is greatly appreciated.
(e-mail: K.Mulcahy at sheffield.ac.uk)
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