contamination in MACS seperated cells - (Mar/31/2008 )
hi all,
I am new to this forum and this is my first post. I am trying to separate monocytes using MACS column but when I culture them further for differentiation to dendritic cells I get conmtamination. can anyone please help me with this? what all precautions should i take while using the column in laminar hood?
desperately waiting for respnse
goofy 17
I am new to this forum and this is my first post. I am trying to separate monocytes using MACS column but when I culture them further for differentiation to dendritic cells I get conmtamination. can anyone please help me with this? what all precautions should i take while using the column in laminar hood?
desperately waiting for respnse
goofy 17
As long as all your buffers, beads, samples have been kept sterile, and you use good sterile techniques you should be fine. Perhaps you should make up your buffers and media again?
I am new to this forum and this is my first post. I am trying to separate monocytes using MACS column but when I culture them further for differentiation to dendritic cells I get conmtamination. can anyone please help me with this? what all precautions should i take while using the column in laminar hood?
desperately waiting for respnse
goofy 17
As long as all your buffers, beads, samples have been kept sterile, and you use good sterile techniques you should be fine. Perhaps you should make up your buffers and media again?
Dear Clare,
thanks a lot for ur reply. actually i have checked my buffers and media. they are fine. could it be that as the mouth of the column is too open, it is inviting contaminants. actually our flow hood is too old and we cant afford to replace it also...can i cover the mouth of the column by autoclaved foil cap so that it is opened only for very short spans?
[/quote]
Dear Clare,
thanks a lot for ur reply. actually i have checked my buffers and media. they are fine. could it be that as the mouth of the column is too open, it is inviting contaminants. actually our flow hood is too old and we cant afford to replace it also...can i cover the mouth of the column by autoclaved foil cap so that it is opened only for very short spans?
[/quote]
Has your hood been serviced recently? You could try covering the column and make sure your cells drip directly into a tube (ie: not from a great height). Have you tried putting some antibiotics in your cells? After FACS sorting I would always put gentamycin in my cells.