A Simple Protocol for the Isolation and Culture of Hepatocytes from MASLD Mice
Peng Jiang1, Zhidan Zhang2, Meijie Chen3
1Department of Endocrinology, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China; Department of Pharmacy, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China.
Abstract:
Metabolic dysfunction-associated steatotic liver disease (MASLD), characterized by pathological lipid accumulation (steatosis) within hepatocytes, represents a significant and growing global health burden. Primary mouse hepatocyte cultures serve as indispensable ex vivo models for elucidating MASLD pathogenesis and therapeutic interventions. However, reliable isolation of high-quality hepatocytes from steatotic livers remains technically challenging. Here, we present a simple protocol for the efficient isolation of primary hepatocytes from MASLD mice using the collagenase perfusion technique. This protocol generates hepatocytes exhibiting inherent steatosis without requiring artificial induction via palmitate and oleic acid supplementation, thereby preserving a more physiologically relevant phenotype. The protocol yields highly viable and high-purity hepatocytes and is universally applicable for isolating high quantities of hepatocytes from MASLD mice. Furthermore, it is suitable for pharmacological evaluation or genetic intervention studies. In summary, we provide a reproducible protocol to isolate high-yield, high-purity, and highly viable hepatocytes for downstream cell biological studies to facilitate the discovery of novel therapeutic targets and drugs for MASLD.
Insights
Researchers developed a simple protocol to isolate high-quality, inherently steatotic primary hepatocytes from metabolic dysfunction-associated steatotic liver disease (MASLD) mouse models. This method aids in studying MASLD pathogenesis and discovering new therapies.
Area of Science:
- Hepatology
- Cell Biology
- Biochemistry
Background:
- Metabolic dysfunction-associated steatotic liver disease (MASLD) is a growing global health concern characterized by liver steatosis.
- Primary mouse hepatocytes are crucial for studying MASLD, but isolating them from steatotic livers is difficult.
- Existing methods often require artificial induction of steatosis, compromising physiological relevance.
Purpose of the Study:
- To present a simple, efficient protocol for isolating primary hepatocytes from MASLD mouse models.
- To obtain hepatocytes with inherent steatosis for more physiologically relevant research.
- To facilitate the study of MASLD pathogenesis and the development of novel therapeutics.
Main Methods:
- Utilized collagenase perfusion technique for hepatocyte isolation from MASLD mouse livers.
- Focused on isolating hepatocytes with inherent steatosis, avoiding artificial induction.
- Emphasized achieving high yield, purity, and viability of isolated hepatocytes.
Main Results:
- Successfully isolated primary hepatocytes exhibiting inherent steatosis from MASLD mice.
- The protocol generated high-yield, high-purity, and highly viable hepatocytes.
- The isolated hepatocytes maintained a physiologically relevant steatotic phenotype.
Conclusions:
- The developed protocol provides a reproducible method for isolating primary hepatocytes from MASLD mice.
- This technique enables downstream cell biological studies, pharmacological evaluations, and genetic intervention studies.
- Facilitates the discovery of novel therapeutic targets and drugs for MASLD.
