《生命科学》 2026, 38(10): 1859-1874

肝细胞分离与培养技术的研究进展及其在畜牧业中的应用

李文微^1,2 , 王云琴^1,2 , 汤少勋³ , 韩雪峰^3,* , 谭支良³ , 龙玲^1,2,*

¹西北民族大学生命科学与工程学院，兰州 730030 ²西北民族大学生物工程学院，兰州 730030 ³中国科学院亚热带农业生态研究所/饲草种质高效设计与利用全国重点实验室/畜禽养殖污染控制与资源化技术国家工程实验室/农业农村部中南动物营养与饲料科学观测实验站/动物营养生理与代谢过程湖南省重点实验室，长沙 410125

摘　要：

肝细胞是执行肝脏代谢、合成、解毒等核心功能的关键细胞，建立能够模拟其体内功能的体外肝细胞模型对研究肝脏生理与病理机制至关重要。经过分离与培养技术的不断优化，肝细胞分离方法从机械法、酶消化法发展至灌流法，显著提高了肝细胞活力和获得率；培养体系则从二维平面培养发展至三维仿生培养。其中，微流控芯片或类器官等三维培养新技术不仅显著提升了肝细胞功能的维持时间，还能高度模拟肝脏复杂微结构与微环境，使其更接近生理状态。体外肝细胞培养为畜牧业研究提供了重要研究平台，在畜禽肝脏生理功能、代谢性疾病机制研究、饲料安全性评估及兽药研发等领域具有重要应用价值。本文系统综述肝细胞分离与培养技术的研究进展，重点分析了各项技术的优势与局限性，并探讨了肝细胞分离与培养技术在畜牧业中的应用。

通讯作者：韩雪峰 , Email:xfhan@isa.ac.cn 龙玲 , Email:longwsm@163.com

Research progress in hepatocyte isolation and culture techniques and their applications in animal husbandry

LI Weng-Wei^1,2 , WANG Yun-Qin^1,2 , TANG Shao-Xun³ , HAN Xue-Feng^3,* , N Zhi-Liang³ , LONG Ling^1,2,*

¹School of Life Science and Engineering, Northwest Minzu University, Lanzhou 730030, China ²College of Bioengineering, Northwest Minzu University, Lanzhou 730030, China ³Institute of Subtropical Agriculture, Chinese Academy of Sciences / National Key Laboratory of Forage Germplasm Innovation and Utilization / National Engineering Laboratory for Pollution Control and Resource Utilization in Livestock and Poultry Production / Central South Scientific Observation and Experiment Station of Animal Nutrition and Feed Science, Ministry of Agriculture and Rural Affairs / Hunan Provincial Key Laboratory of Animal Nutritional Physiology and Metabolic Process, Changsha 410125, China

Abstract:

Hepatocytes are the key functional cells responsible for hepatic metabolism, synthesis, and detoxification, playing an irreplaceable role in maintaining liver homeostasis. The in vivo microenvironment of hepatocytes is intricate, involving interactions with non-parenchymal cells, extracellular matrix, and hemodynamic signals, which are essential for sustaining their normal physiological functions. Establishing in vitro hepatocyte models that can accurately simulate their in vivo functions is crucial for studying liver physiology and pathological mechanisms, as well as providing technical support for related research in biomedicine and animal husbandry. Through continuous optimization of isolation and culture techniques, the isolation methods of hepatocytes have evolved from mechanical dissociation and enzymatic digestion to perfusion. Mechanical dissociation is simple but causes severe cell damage, while enzymatic digestion improves yield but has uneven effects; perfusion, by contrast, minimizes cell damage while fully digesting liver tissue, significantly improving hepatocyte viability and yield. Meanwhile, culture systems have advanced from two-dimensional (2D) monolayer cultures to three dimensional (3D) biomimetic models. 2D monolayer culture is easy to operate and low-cost but fails to maintain long-term hepatocyte functions, with obvious differences from in vivo states. Notably, emerging 3D culture technologies—such as microfluidic chips and organoids—not only prolong the functional maintenance of hepatocytes but also highly recapitulate the liver's complex microstructure and microenvironment, closely mimicking physiological conditions. In vitro hepatocyte culture also serves as a vital research platform for animal husbandry, with significant applications in livestock liver physiology, metabolic disease mechanisms, feed safety evaluation, and veterinary drug development. This review systematically summarizes recent advances in hepatocyte isolation and culture techniques, analyzes their respective advantages and limitations, and discusses their practical applications and development prospects in animal husbandry, providing a comprehensive reference for subsequent related research.

Communication Author：HAN Xue-Feng , Email:xfhan@isa.ac.cn LONG Ling , Email:longwsm@163.com