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2016 ; 11
(3
): e0151945
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Methods for Isolation and Purification of Murine Liver Sinusoidal Endothelial
Cells: A Systematic Review
#MMPMID26992171
Meyer J
; Gonelle-Gispert C
; Morel P
; Bühler L
PLoS One
2016[]; 11
(3
): e0151945
PMID26992171
show ga
To study the biological functions of liver sinusoidal endothelial cells (LSEC)
and to identify their interplay with blood or liver cells, techniques allowing
for the isolation and purification of LSEC have been developed over the last
decades. The objective of the present review is to summarize and to compare the
efficiency of existing methods for isolating murine LSEC. Toward this end, the
MEDLINE database was searched for all original articles describing LSEC isolation
from rat and mouse livers. Out of the 489 publications identified, 23 reported
the main steps and outcomes of the procedure and were included in our review.
Here, we report and analyse the technical details of the essential steps of the
techniques used for LSEC isolation. The correlations between the prevalence of
some steps and the efficiency of LSEC isolation were also identified. We found
that centrifugal elutriation, selective adherence and, more recently,
magnetic-activated cell sorting were used for LSEC purification. Centrifugal
elutriation procured high yields of pure LSEC (for rats 30-141.9 million cells
for 85-98% purities; for mice 9-9.25 million cells for >95% purities), but the
use of this method remained limited due to its high technical requirements.
Selective adherence showed inconsistent results in terms of cell yields and
purities in rats (5-100 million cells for 73.7-95% purities). In contrast,
magnetic-activated cell sorting allowed for the isolation of highly pure LSEC,
but overall lower cell yields were reported (for rats 10.7 million cells with
97.6% purity; for mice 0.5-9 million cells with 90-98% purities). Notably, the
controversies regarding the accuracy of several phenotypic markers for LSEC
should be considered and their use for both magnetic sorting and characterization
remain doubtful. It appears that more effort is needed to refine and standardize
the procedure for LSEC isolation, with a focus on the identification of specific
antigens. Such a procedure is required to identify the molecular mechanisms
regulating the function of LSEC and to improve our understanding of their role in
complex cellular processes in the liver.
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