Please share the method to follow, immunomagnetic sorting or flowcytometry ?
How to get rid of other cell population apart from blast cells?
What should be the culture medium and how does culture behave?
How long can we maintain such cultures?
We used to isolate CD34 positive cells by magnetic microbeads through direct staining with anti-CD34.
Then, you are able to isolate these cells and get rid of the rest. We usually get a 98,5% of purenes or higher with this method.
You can get easily the protocol on milteny website.
Hope it has helped!
But not all AML subtypes express CD34. So in these cases you need to sort them based on other markers expression (usually it might be CD117 - but more problematic would be cases of AML-M5).
Flowcytomertry sorting can provided more purity of cell by using multi-marker sort. Usually magnetic beads sort is operated easily. You have your choice between the two methods depended on your main purpose of your further study after sorting.
I agree for AML-M5 (monoblastic). CD64, CD11b and lysozyme are characteristically positive in AMoL. CD33 and CD64 are consistently expressed in 100% and 96% of vcases, respectively (CD33 is generally bright).
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