Open AccessAn Improved Time- and Labor- Efficient Protocol for Mouse Primary Hepatocyte Isolation
Author(s) -
Mingxiao Feng,
Sara A. DiVall,
Sheng Wu
Publication year - 2021
Publication title -
journal of visualized experiments
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.596
H-Index - 91
ISSN - 1940-087X
DOI - 10.3791/61812
Subject(s) - hepatocyte , protocol (science) , isolation (microbiology) , primary (astronomy) , in vitro , computational biology , computer science , biology , chemistry , bioinformatics , biochemistry , medicine , pathology , physics , alternative medicine , astronomy
Primary hepatocytes are used extensively in liver in vitro research, especially in glucose metabolism studies. A base technique has been adapted based on different needs, like time, labor, cost, and primary hepatocyte usage, resulting in various primary hepatocyte isolation protocols. However, the numerous steps and time-consuming reagent preparations in primary hepatocyte isolation are major drawbacks for efficiency. After comparing different protocols for their pros and cons, the advantages of each were combined, and a rapid and efficient primary hepatocyte isolation protocol was formulated. Within only ~35 min, this protocol could yield as much, if not more, healthy primary hepatocytes as other protocols. Further, glucose metabolism experiments performed using the isolated primary hepatocytes validated the usefulness of this protocol in in vitro liver metabolism studies. We also extensively reviewed and analyzed the significance and purpose of each step in this study so that future researchers can further optimize this protocol based on needs.