Medizinische Universität Graz - Research portal
Selected Publication:
Passegger, CA.
Relative enrichment and analysis of rare cells from blood.
[ Diplomarbeit/Master Thesis ] University of Graz and University of Technology Graz; 2013. pp.74.
- Authors Med Uni Graz:
- Advisor:
- Kroneis Thomas
- Sedlmayr Peter
- Altmetrics:
- Abstract:
- Invasive prenatal diagnosis techniques such as chorionic villus sampling, amniocentesis and cordocentesis increase the risk of fetal loss and entail the possibility of other procedure-related complications. Therefore non-invasive approaches based on free fetal cells or cell-free fetal DNA circulating in the maternal blood are important areas of research. Fetal cells as well as for example circulating tumor cells belong to the group of rare cells. As the name implies these cells occur in a very low frequency with an incidence of approximately one cell per ml maternal blood. That is why enrichment is a necessary, but at the same time a challenging task. In this study three methods for the relative enrichment of rare cells from peripheral blood were compared. The two methods tested that use immunomagnetic particles for the depletion of CD45+ cells led to rather modest results. Recovery rates achieved with these approaches did not exceed 22%. In contrast, the isolation of the target cells by size with a filter achieved results with an average recovery of approximately 89%. This method was then used to evaluate DNA quality. With 2% formaldehyde treated, filtrated and hematoxylin stained cells were isolated with laser capture microdissection for subsequent analysis. Whole genome amplification was successful in 2/3 of the analyzed cells. In addition, 50% of the overall samples analyzed also had good DNA quality.