Related Experiment Video
Updated: Apr 4, 2026
06:32
Isolation, Enrichment, and Maintenance of Medulloblastoma Stem Cells
Published on: September 1, 2010
16.9K
Enriched Bone Marrow Derived Disseminated Neuroblastoma Cells Can Be a Reliable Source for Gene Expression Studies-A
Fikret Rifatbegovic1, M Reza Abbasi1, Sabine Taschner-Mandl1
1CCRI, Children's Cancer Research Institute, St. Anna Kinderkrebsforschung, Vienna, Austria.
Plos One
|September 12, 2015
Summary
Pre-analytical processing of bone marrow samples for disseminated tumor cells (DTCs) is crucial. While magnetic bead enrichment is safe for gene expression, transport temperature and duration significantly impact results in neuroblastoma patients.
Area of Science:
- Oncology
- Molecular Biology
- Biotechnology
Background:
- Disseminated tumor cells (DTCs) in bone marrow (BM) are common in high-risk neuroblastoma.
- Enrichment strategies are needed for DTC characterization due to their low frequency.
- Pre-analytical processing steps may introduce artifacts in gene expression analysis.
Purpose of the Study:
- To evaluate the impact of pre-analytical processing on gene expression in DTCs.
- To determine if tumor cell enrichment techniques affect gene expression profiles.
- To assess the influence of transport duration and temperature on DTC gene expression.
Main Methods:
- Neuroblastoma cells (LAN-1) were spiked into tumor-free BM and peripheral blood.
- Samples were subjected to various storage conditions (room temperature, 4°C, -80°C) for different durations.
- Tumor cells were enriched using magnetic beads, and gene expression was analyzed via qPCR and microarrays.
Main Results:
- Storage at -80°C (with DMSO) and subsequent thawing did not significantly alter gene expression.
- Magnetic bead enrichment of neuroblastoma cells showed no significant effect on gene expression.
- Storage at 4°C affected gene expression at 72 hours, while room temperature storage impacted it significantly from 24 hours onwards.
Conclusions:
- Magnetic bead separation is a reliable method for enriching DTCs without compromising gene expression analysis.
- Transport time and temperature are critical factors that can significantly alter DTC gene expression profiles.
- Routinely collected BM samples can be analyzed for gene expression, provided transport conditions are carefully monitored.
