Evidence map›Paper›PMID 27077489›Full record

ArticleJournal of visualized experiments : JoVE2016

Protocol for Isolation of Primary Human Hepatocytes and Corresponding Major Populations of Non-parenchymal Liver Cells.

Victoria Kegel, Daniela Deharde, Elisa Pfeiffer, Katrin Zeilinger, Daniel Seehofer, Georg Damm

Open access · bronzeAbstract readVideo-Audio Media
In one paragraph

Article in Journal of visualized experiments : JoVE, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 64 papers.

0numbers the graph read from it
0cells of the map it votes in
64citing papers in PubMed
5.1field-weighted citation impact, top 4% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

64 citing papers in PubMed, 96 citations in OpenAlex.

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  7. Ex vivo Drug Metabolism Using Primary Human Hepatocytes for the Detection of Metabolite-Dependent Antibodies to Red Blood Cells.Transfusion medicine and hemotherapy : offizielles Organ der Deutschen Gesellschaft fur Transfusionsmedizin und Immunhamatologie · 2025
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4 more citing papers are in PubMed but not listed here.

4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

6 authors at 2 institutions in 1 country.

Victoria KegelDepartment of General-, Visceral- and Transplantation Surgery, Charité University Medicine Berlin.
Daniela DehardeDepartment of General-, Visceral- and Transplantation Surgery, Charité University Medicine Berlin.
Elisa PfeifferDepartment of General-, Visceral- and Transplantation Surgery, Charité University Medicine Berlin.
Katrin ZeilingerBrandenburg Center for Regenerative Therapies, Charité University Medicine Berlin.
Daniel SeehoferDepartment of General-, Visceral- and Transplantation Surgery, Charité University Medicine Berlin.
Georg DammDepartment of General-, Visceral- and Transplantation Surgery, Charité University Medicine Berlin; Georg.Damm@charite.de.
Charité - Universitätsmedizin Berlin · DEBerlin-Brandenburger Centrum für Regenerative Therapien · DE

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Beside parenchymal hepatocytes, the liver consists of non-parenchymal cells (NPC) namely Kupffer cells (KC), liver endothelial cells (LEC) and hepatic Stellate cells (HSC). Two-dimensional (2D) culture of primary human hepatocyte (PHH) is still considered as the "gold standard" for in vitro testing of drug metabolism and hepatotoxicity. It is well-known that the 2D monoculture of PHH suffers from dedifferentiation and loss of function. Recently it was shown that hepatic NPC play a central role in liver (patho-) physiology and the maintenance of PHH functions. Current research focuses on the reconstruction of in vivo tissue architecture by 3D- and co-culture models to overcome the limitations of 2D monocultures. Previously we published a method to isolate human liver cells and investigated the suitability of these cells for their use in cell cultures in Experimental Biology and Medicine(1). Based on the broad interest in this technique the aim of this article was to provide a more detailed protocol for the liver cell isolation process including a video, which will allow an easy reproduction of this technique. Human liver cells were isolated from human liver tissue samples of surgical interventions by a two-step EGTA/collagenase P perfusion technique. PHH were separated from the NPC by an initial centrifugation at 50 x g. Density gradient centrifugation steps were used for removal of dead cells. Individual liver cell populations were isolated from the enriched NPC fraction using specific cell properties and cell sorting procedures. Beside the PHH isolation we were able to separate KC, LEC and HSC for further cultivation. Taken together, the presented protocol allows the isolation of PHH and NPC in high quality and quantity from one donor tissue sample. The access to purified liver cell populations could allow the creation of in vivo like human liver models.

Indexed as

Cell SeparationCoculture TechniquesEndothelial CellsHepatic Stellate CellsHepatocytesHumansKupffer CellsLiver

Identifiers

PMID27077489
PMCPMC4841324
OpenAlexW2312716565

What OpenQuestion holds

Textmetadata
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.