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Hematopoietic cells separation


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#1 soosin

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Posted 11 January 2011 - 06:21 AM

I have been testing MACS separation column for sometime, but still can't get good yield. Altough i tried to increase the volume of blood in order to get high lymphocyte count as high as 13 million cells, but still facing problem in isolate them. My cells of interest are CD4+, CD8+ and CD56+.
Can anyone here suggest me the steps to separate those cells efficiently? -Thanks-

Regards,
Soosin

#2 marco77

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Posted 12 January 2011 - 02:08 PM

I have been testing MACS separation column for sometime, but still can't get good yield. Altough i tried to increase the volume of blood in order to get high lymphocyte count as high as 13 million cells, but still facing problem in isolate them. My cells of interest are CD4+, CD8+ and CD56+.
Can anyone here suggest me the steps to separate those cells efficiently? -Thanks-

Regards,
Soosin


Hi, I am not sure but you could tried to use ficoll with different density, maybe these cells are a particular density, try to use 1,077 or 1,090 mixed with RPMI or fisiological condition to obtain different density, its a STRANGE IDEA BUT sometime something happen!
Good luck
Marco

#3 soosin

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Posted 13 January 2011 - 02:56 AM


I have been testing MACS separation column for sometime, but still can't get good yield. Altough i tried to increase the volume of blood in order to get high lymphocyte count as high as 13 million cells, but still facing problem in isolate them. My cells of interest are CD4+, CD8+ and CD56+.
Can anyone here suggest me the steps to separate those cells efficiently? -Thanks-

Regards,
Soosin


Hi, I am not sure but you could tried to use ficoll with different density, maybe these cells are a particular density, try to use 1,077 or 1,090 mixed with RPMI or fisiological condition to obtain different density, its a STRANGE IDEA BUT sometime something happen!
Good luck
Marco




Thanks for answering my question.
I have stained the cell populations before cell separation and confirm that they are all isolated from blood. Just not sure if they are very little.
Your answer could be helpful to me, i will try it out.
Thanks.




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