Publication: Quantitative mRNA expression analysis in
kidney glomeruli using microdissection techniques
Authors
De Spiegelaere, Ward ; Cornillie, Pieter ; Van Poucke, Mario ; Peelman, Luc ; Burvenich, Christian ; Van den Broeck, Wim
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Publisher
Murcia: F. Hernández
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DOI
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info:eu-repo/semantics/article
Description
Abstract
The introduction of new tools for molecular
analysis, such as RT-qPCR and microarrays, has
provided researchers with powerful applications to study
renal disease and development. However, the high
cellular heterogeneity of the renal tissue complicates the
molecular analysis of specific cells and cell groups such
as glomerular or tubular cells. In the past, glomerular
sieving and manual dissection were used for the isolation
of glomeruli. However, these techniques cannot be used
for the isolation of specific glomeruli or for the coisolation
of additional tissue fractions. In recent decades,
new microdissection techniques such as laser-assisted
microdissection have been developed. These
applications allow the isolation of small cell groups from
heterogeneous tissue for molecular analysis, including
microarray and RT-qPCR. Although very promising,
some drawbacks are associated with these techniques.
The isolated sample material is generally small and
requires sensitive assays. In addition, the long sample
processing time may result in a considerable loss of
RNA integrity. Careful optimization and rigorous quality
analysis should overcome these drawbacks. In the
present paper, the recent literature on the application of
microdissection techniques in kidney research is
reviewed, together with a discussion of the critical issues
that are essential for the application of quantitative
mRNA expression analysis with RT-qPCR on
microdissected samples.
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