Abstract：
:The objective of the present study was to develop an accurate and reproducible method for liver sinusoidal endothelial cell (LSEC) isolation in mice. Non-parenchymal cells were isolated using a modified two-step collagenase digestion combined with Optiprep density gradient centrifugation. LSEC were further purified using two prevalent methods, short-term selective adherence and CD146+ magnetic-activated cell sorting (MACS), and compared in terms of cell yield, viability and purity to our purification technique using CD11b cell depletion combined with long-term selective adherence. LSEC purification using our technique allowed to obtain 7.07±3.80 million LSEC per liver, while CD146+ MACS and short-term selective adherence yielded 2.94±1.28 and 0.99±0.66 million LSEC, respectively. Purity of the final cell preparation reached 95.10±2.58% when using our method. In contrast, CD146+ MACS and short-term selective adherence gave purities of 86.75±3.26% and 47.95±9.82%, respectively. Similarly, contamination by non-LSEC was the lowest when purification was performed using our technique, with a proportion of contaminating macrophages of only 1.87±0.77%. Further, isolated cells analysed by scanning electron microscopy presented typical LSEC fenestrations organized in sieve plates, demonstrating that the technique allowed to isolate bona fide LSEC. In conclusion, we described a reliable and reproducible technique for the isolation of high yields of pure LSEC in mice. This protocol provides an efficient method to prepare LSEC for studying their biological functions.
journal_name
Exp Cell Resjournal_title
Experimental cell researchauthors
Meyer J,Lacotte S,Morel P,Gonelle-Gispert C,Bühler Ldoi
10.1016/j.yexcr.2016.10.024subject
Has Abstractpub_date
2016-12-10 00:00:00pages
291-301issue
2eissn
0014-4827issn
1090-2422pii
S0014-4827(16)30359-7journal_volume
349pub_type
杂志文章abstract：:Endothelial cells are exposed to fluid shear stress profiles that vary in magnitude, pulsatility, and directionality due to regional variations in blood vessel structure. Laminar flow at physiological levels is atheroprotective; multidirectional or reversing low (disturbed) flow promotes inflammation and disease; and ...
journal_title：Experimental cell research
pub_type： 杂志文章,评审
doi：10.1016/j.yexcr.2019.01.005
更新日期：2019-03-01 00:00:00
abstract：OBJECTIVES:Our purpose was to investigate the role of the nitric oxide (NO) signaling pathway in the regulation of ciliary beat frequency (CBF) in mouse nasal and tracheal epithelial cells. METHODS:We studied the effects of the NO donor l-arginine (L-Arg) and specific inhibitors of the NO signaling pathway on CBF of b...
journal_title：Experimental cell research
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doi：10.1016/j.yexcr.2011.07.007
更新日期：2011-10-15 00:00:00
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journal_title：Experimental cell research
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doi：10.1016/0014-4827(91)90264-u
更新日期：1991-10-01 00:00:00
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更新日期：2016-08-01 00:00:00
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journal_title：Experimental cell research
pub_type： 杂志文章
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journal_title：Experimental cell research
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journal_title：Experimental cell research
pub_type： 杂志文章
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更新日期：1995-10-01 00:00:00
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pub_type： 杂志文章
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journal_title：Experimental cell research
pub_type： 杂志文章
doi：10.1016/j.yexcr.2018.05.037
更新日期：2018-08-15 00:00:00
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journal_title：Experimental cell research
pub_type： 杂志文章,评审
doi：10.1006/excr.1999.4668
更新日期：1999-11-25 00:00:00
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journal_title：Experimental cell research
pub_type： 杂志文章
doi：10.1016/j.yexcr.2020.112402
更新日期：2021-01-01 00:00:00
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journal_title：Experimental cell research
pub_type： 杂志文章
doi：10.1006/excr.1996.0159
更新日期：1996-05-25 00:00:00
abstract：:In the present study, we have tested the influence of inhibition of endocytosis by hypertonic medium on the regulation of cell surface insulin receptors. We show that active internalization of 125I-insulin is markedly inhibited by hypertonic media and that, in parallel, cell surface invaginations are significantly dim...
journal_title：Experimental cell research
pub_type： 杂志文章
doi：10.1016/0014-4827(89)90034-7
更新日期：1989-11-01 00:00:00
abstract：:LMX1B is a LIM-homeodomain transcription factor essential for development. Putative LMX1B target genes have been identified through mouse gene targeting studies, but their identity as direct LMX1B targets remains hypothetical. We describe here the first molecular characterization of LMX1B target gene regulation. Micro...
journal_title：Experimental cell research
pub_type： 杂志文章
doi：10.1016/j.yexcr.2008.10.012
更新日期：2009-01-01 00:00:00
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journal_title：Experimental cell research
pub_type： 杂志文章
doi：10.1016/j.yexcr.2014.02.028
更新日期：2014-05-01 00:00:00
abstract：:Breast cancer is the most common carcinoma that metastasizes to bone. Tumor-produced parathyroid hormone-related protein (PTHrP), a known stimulator of osteoclastic bone resorption, is a major mediator of the osteolytic process in breast cancer. We have previously shown that PTHrP increases breast cancer cell prolifer...
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pub_type： 杂志文章
doi：10.1016/j.yexcr.2006.08.011
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journal_title：Experimental cell research
pub_type： 杂志文章
doi：10.1016/j.yexcr.2010.08.011
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pub_type： 杂志文章
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更新日期：2011-06-10 00:00:00
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pub_type： 杂志文章
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pub_type： 杂志文章
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更新日期：2016-09-10 00:00:00
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pub_type： 杂志文章
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journal_title：Experimental cell research
pub_type： 杂志文章
doi：10.1006/excr.1997.3728
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pub_type： 杂志文章
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更新日期：1984-03-01 00:00:00
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journal_title：Experimental cell research
pub_type： 杂志文章
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更新日期：1985-12-01 00:00:00
abstract：:USF is a small family of basic helix-loop-helix leucine zipper (bHLH-zip) transcription factors with DNA binding specificities similar to that of the c-Myc oncoprotein. Evidence for a role of USF in growth control includes inhibition of c-Myc-dependent cellular transformation in vitro and loss of USF transcriptional a...
journal_title：Experimental cell research
pub_type： 杂志文章
doi：10.1016/j.yexcr.2004.08.013
更新日期：2005-01-01 00:00:00
abstract：:Coiled bodies (CBs) are structural constituents observed in nuclei of most eukaryotic cells. They usually occur in the nucleoplasm as well as in contact with the nucleolar surface. In this work we studied the hepatocyte nuclei of hibernating dormice in order to investigate possible modifications of CBs along the seaso...
journal_title：Experimental cell research
pub_type： 杂志文章
doi：10.1006/excr.1994.1106
更新日期：1994-04-01 00:00:00
