[ Title ]
- Toward Rare Blood Cell Preservation for RNA Sequencing
[ Journal ]
- JOURNAL OF MOLECULAR DIAGNOSTICS
[ Author ]
- Vickovic, S
Ahmadian, A
Lewensohn, R
Lundeberg, J
[ Abstract ]
- Cancer is driven by various events Leading to cell differentiation and
disease progression. Molecular tools are powerful approaches for
describing how and why these events occur. With the growing field of
next-generation DNA sequencing, there is an increasing need for
high-quality nucleic acids derived from human cells and tissues a
prerequisite for successful cell profiting. Although advances in RNA
preservation have been made, some of the largest biobanks still do not
employ RNA blood preservation as standard because of Limitations in low
blood-input volume and RNA stability over the whole gene body.
Therefore, we have developed a robust protocol for blood preservation
and tong-term storage while maintaining RNA integrity. Furthermore, we
explored the possibility of using the protocol for preserving rare cell
samples, such as circulating tumor cells. The results of our study
confirmed that gene expression was not impacted by the preservation
procedure (r(2) > 0.88) or by Long-term storage (r(2) = 0.95), with RNA
integrity number values averaging over 8. Similarly, cell surface
antigens were still available for antibody selection (r(2) = 0.95).
Lastly, data mining for fusion events showed that it was possible to
detect rare tumor cells among a background of other cells present in
blood irrespective of fixation. Thus, the developed protocol would be
suitable for rare blood cell preservation followed by RNA sequencing
analysis.
[ URL ]
- http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=Alerting&SrcApp=Alerting&DestApp=WOS&DestLinkType=FullRecord;UT=WOS:000357441600003