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We aimed at developing a new antibody\u2010free isolation method able to obtain the four main hepatic cell types (hepatocytes, liver sinusoidal endothelial cells [\n                    <jats:styled-content style=\"fixed-case\">LSEC<\/jats:styled-content>\n                    ], hepatic macrophages [\n                    <jats:styled-content style=\"fixed-case\">HM<\/jats:styled-content>\n                    \u03a6] and hepatic stellate cells [\n                    <jats:styled-content style=\"fixed-case\">HSC<\/jats:styled-content>\n                    ]) from a single rat liver. Control and cirrhotic (\n                    <jats:styled-content style=\"fixed-case\">CC<\/jats:styled-content>\n                    l\n                    <jats:sub>4<\/jats:sub>\n                    and\n                    <jats:styled-content style=\"fixed-case\">TAA<\/jats:styled-content>\n                    ) rat livers (n\u00a0=\u00a06) were perfused, digested with collagenase and mechanically disaggregated obtaining a multicellular suspension. Hepatocytes were purified by low revolution centrifugations while non\u2010parenchymal cells were subjected to differential centrifugation. Two different fractions were obtained:\n                    <jats:styled-content style=\"fixed-case\">HSC<\/jats:styled-content>\n                    and mixed\n                    <jats:styled-content style=\"fixed-case\">LSEC<\/jats:styled-content>\n                    +\n                    <jats:styled-content style=\"fixed-case\">HM<\/jats:styled-content>\n                    \u03a6. Further\n                    <jats:styled-content style=\"fixed-case\">LSEC<\/jats:styled-content>\n                    and\n                    <jats:styled-content style=\"fixed-case\">HM<\/jats:styled-content>\n                    \u03a6 enrichment was achieved by selective adherence time to collagen\u2010coated substrates. Isolated cells showed high viability (80%\u201095%) and purity (&gt;95%) and were characterized as functional: hepatocytes synthetized albumin and urea,\n                    <jats:styled-content style=\"fixed-case\">LSEC<\/jats:styled-content>\n                    maintained endocytic capacity and in\u00a0vivo fenestrae distribution,\n                    <jats:styled-content style=\"fixed-case\">HM<\/jats:styled-content>\n                    \u03a6 increased expression of inflammatory markers in response to\n                    <jats:styled-content style=\"fixed-case\">LPS<\/jats:styled-content>\n                    and\n                    <jats:styled-content style=\"fixed-case\">HSC<\/jats:styled-content>\n                    were activated upon in\u00a0vitro culture. The 4 in 1 protocol allows the simultaneous isolation of highly pure and functional hepatic cell sub\u2010populations from control or cirrhotic single livers without antibody selection.\n                  <\/jats:p>","DOI":"10.1111\/jcmm.13988","type":"journal-article","created":{"date-parts":[[2018,11,12]],"date-time":"2018-11-12T03:26:54Z","timestamp":1541993214000},"page":"877-886","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":18,"title":["4 in 1: Antibody\u2010free protocol for isolating the main hepatic cells from healthy and cirrhotic single rat livers"],"prefix":"10.1111","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2364-6675","authenticated-orcid":false,"given":"Anabel","family":"Fern\u00e1ndez\u2010Iglesias","sequence":"first","affiliation":[{"name":"Liver Vascular Biology Research Group Barcelona Hepatic Hemodynamic Laboratory IDIBAPS Biomedical Research Institute Barcelona 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