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Purification of Hepatocyte and Sinusoidal Endothelial Cells from Mouse Liver Perfusion
Author(s) -
Cabral Fatima,
Miller Colton M.,
Kudrna Katrina M.,
Hass Blake E.,
Daubendiek Jocelyn G.,
Keller Briana M.,
Harris Edward N.
Publication year - 2018
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2018.32.1_supplement.541.6
Subject(s) - microbiology and biotechnology , endothelial stem cell , hepatocyte , liver cytology , cell , biology , cell type , in vivo , chemistry , in vitro , biochemistry , liver metabolism
The liver is primarily composed of hepatocytes and the nonparenchymal cells which include stellate cells, Kupffer cells, sinusoidal endothelial cells and various other cell populations in very low numbers such as transient leukocytes. Cell isolation with high purity is required for many experiments related to the specific biology and biochemistry of these very different cell types. Here, we outline a relatively simple procedure to obtain a high yield of live hepatocytes and sinusoidal endothelial cells. Additionally, this procedure is relatively simple and does not require specialized equipment other than a refrigerated table top centrifuge and peristatic pump. The mouse portal vein is used as the catheterization site rather than the vena cava, limiting contamination of other cell types during the isolation process. Access to the vein is quick minimizing ischemia and reperfusion of the liver which reduces hepatic cell viability. Dead hepatocytes are also easily identified during centrifugation steps due to differences in cell density. This process allows for isolation of hepatocytes with 99% cell purity and isolation of sinusoidal endothelial cells with approximately 89.1% cell purity. This method will help to answer key questions in the hepatotology field such as determining specific cellular functions with in vitro assays or assessing individual cells after an in vivo treatment. Support or Funding Information Funding is provided in part by the NIH from grant R01HL130864. This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .