Scientific article

An optimized method for mouse liver sinusoidal endothelial cell isolation

Published inExperimental cell research, vol. 349, no. 2, p. 291-301
Publication date2016

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.

  • CD11b
  • LSEC
  • Mice
  • Purification
  • Stabilin-2
Citation (ISO format)
MEYER, Jérémy et al. An optimized method for mouse liver sinusoidal endothelial cell isolation. In: Experimental cell research, 2016, vol. 349, n° 2, p. 291–301. doi: 10.1016/j.yexcr.2016.10.024
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Article (Published version)
ISSN of the journal0014-4827

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