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Protocol for MicroRNA Transfer into Adult Bone Marrow-derived Hematopoietic Stem Cells to Enable Cell Engineering Combined with Magnetic Targeting
Published on: June 18, 2018
Preanalytical mRNA stabilization of whole bone marrow samples
Claudia Langebrake1, Kalle Günther, Jürgen Lauber
1Department of Pediatric Hematology and Oncology, Hannover Medical School, Hannover, Germany. claudia@langebrake.de
Clinical Chemistry
|February 10, 2007
Summary
The PAXgene Bone Marrow RNA System effectively stabilizes RNA from clinical bone marrow samples, ensuring high-quality RNA for gene expression analysis and reducing sample degradation during storage.
Area of Science:
- Molecular Biology
- Genomics
- Biotechnology
Background:
- Gene expression profiling is crucial for cancer diagnosis and research.
- Short-term storage and RNA isolation of blood or bone marrow (BM) specimens can alter gene expression patterns due to induction, repression, and RNA degradation.
Purpose of the Study:
- To evaluate the effectiveness of the PAXgene Bone Marrow RNA System for stabilizing RNA from BM specimens over time.
- To assess the impact of RNA stabilization on gene expression integrity and RNA isolation success rates.
Main Methods:
- Investigated the PAXgene Bone Marrow RNA System using 256 RNA samples from 64 BM specimens.
- Compared RNA yield, integrity, and gene expression stability between PAXgene-stabilized samples and unstabilized reference samples after storage.
Main Results:
- The PAXgene system demonstrated significantly higher RNA integrity compared to the reference method (P <0.01).
- Stabilized samples showed high pairwise correlation in gene expression after 48-h storage, unlike reference samples which exhibited major differences.
- Unstabilized samples had an approximately 3 times higher failure rate in RNA preparation.
Conclusions:
- The PAXgene system effectively stabilizes RNA from clinical bone marrow samples.
- The system is suitable for isolating high-quality and high-quantity RNA, preserving gene expression patterns for downstream analysis.
