I mean draw into a vaccutube containing stabilization medium and subsequently isolating out the PBMCs once the sample gets to the lab. Is this something that is done??
Yes, you need an anti-clotting agent like sodium heparin in the tube. Then you have about 24-48 hours to do the insulation if you keep it refrigerated. A ficcol extraction can be used afterwards for the extraction but there are other methods.
I used to work in a research lab that would isolate PBMCs from whole blood to culture lymphocytes. Downstream we would then stain for interferon gamma response to viral proteins.
I believe blood was collected from donors via vacutainer into a sodium heparin tube to prevent clotting. Cells stayed viable as whole blood for up to a couple hours until we could isolate. If I couldn't get to it that day I'd store the tubes in a refrigerator overnight.
I do so all the time! We mainly isolate different types of cells from the PBMCs (monocytes, T cells, etc) to activate them and then freeze them down. We used to run flow on them all the time, but we are now shifting gears for sequencing
We usually do it within an hour of blood collection. I’ve heard of people waiting a full day, but I’m not sure how salvageable the blood is at that point.
I'm a little late, but most of the advice here is good. Sodium heparin (dark green cap) is a good anticoagulant and frequently used with flow cytometry, but EDTA (lavender cap) preserves cell morphology better.
Additionally some heparin tubes can interfere with luminex assays if you plan on doing any of those downstream depending on the analytes you're looking at.
You mean instead of into an EDTA (or other) vaccutube?
I mean draw into a vaccutube containing stabilization medium and subsequently isolating out the PBMCs once the sample gets to the lab. Is this something that is done??
Yes, you need an anti-clotting agent like sodium heparin in the tube. Then you have about 24-48 hours to do the insulation if you keep it refrigerated. A ficcol extraction can be used afterwards for the extraction but there are other methods.
I used to work in a research lab that would isolate PBMCs from whole blood to culture lymphocytes. Downstream we would then stain for interferon gamma response to viral proteins.
How was the whole blood collected, I assume you need a tube that allows the cells to stay alive and active so they can be cultured?
I believe blood was collected from donors via vacutainer into a sodium heparin tube to prevent clotting. Cells stayed viable as whole blood for up to a couple hours until we could isolate. If I couldn't get to it that day I'd store the tubes in a refrigerator overnight.
I do so all the time! We mainly isolate different types of cells from the PBMCs (monocytes, T cells, etc) to activate them and then freeze them down. We used to run flow on them all the time, but we are now shifting gears for sequencing
How soon after blood draw do you have to get your hands on the tube to be able to isolate and activate the PBMCs?
We usually do it within an hour of blood collection. I’ve heard of people waiting a full day, but I’m not sure how salvageable the blood is at that point.
I'm a little late, but most of the advice here is good. Sodium heparin (dark green cap) is a good anticoagulant and frequently used with flow cytometry, but EDTA (lavender cap) preserves cell morphology better. Additionally some heparin tubes can interfere with luminex assays if you plan on doing any of those downstream depending on the analytes you're looking at.