Can I preserve whole blood sample with RNAlater and then separate CD14+ cells - by Ficoll centrifugation and incubation at 37*C? (Oct/06/2005 )
I have a technical question regarding RNAlater using in order to preserve PBMC in the whole blood specimen. I have found on Ambion website that by using RNAlater and the RiboPure-Blood kit I can preserve PBMC from the whole blood. Unfortunately, I need only CD14+ cells and my question is following: Is it possible to treat the whole blood with RNA later solution, then store it for recommended time period at the recommended temperature and after that perform Ficoll Paque gradient centrifugation to separate PBMCs, wash these PBMCs and incubate them on 37*C to allow the CD14+ monocytes to attach ~ to separate CD14+ from the rest of the cells? In short, are the cells after RNA later treatment alive? Do they behave as "fresh" cells and keep the ability to attach?
I have to solve this “RNA stabilization” step because my specimens are being collected from several outpatient clinics and it takes time for them to reach our lab to be processed…
Thank you very much for anu suggestion.
I dont think so... I doubt they could survive the precipitation of the proteins in the cell by RNAlater... In my hands it seemed like the primary human T-cells I was using were lysed in the RNAlater so that I couldn't even pellet the cells to remove the RNAlater for downstream applications...
And is there any other way how to preserve RNA in monocytes when impossible to process the blood specimen immediately after blood taking?