Researchers developed novel procedure for isolating primary mouse hepatocytes with holographic acoustic tweezers

Recently, a researcher team led by Prof. LI Fei and ZHENG Hairong from the Shenzhen Institute of Advanced Technology (SIAT) of Chinese Academy of Sciences, along with Prof. MAO Yilei and YANG Huayu from the Peking Union Medical College (PUMC) Hospital, have established a comprehensive procedure for isolating primary mouse hepatocytes and maintaining them in long-term culture with significant amplification in a two-dimensional (2D) environment.

The study was published in Biomaterials on Jul. 01.

Primary cells sourced directly from organisms have become indispensable tools for cellular and molecular biology research in recent years. However, the current three-dimensional (3D) culture models suffer from drawbacks such as a limited lifespan, stringent culture requirements, complicated procedures, and restricted cell numbers, hindering their applications, especially in isolating and culturing primary hepatocytes in vitro.

Holographic acoustic tweezers (HAT) exhibit significant potential in constructing in vitro 3D models for investigating the mechanisms of cancer, testing and selecting suitable anticancer drugs, and developing new medications. However, there is a lack of in-depth research on the biological effects of HAT on primary cells.

In this study, the researcher improved the existing HAT technology and proposed "Holographic lattice-based acoustic tweezers". They developed a group of acoustic holographic lenses to modulate the phase and amplitude of the incident sound wave simultaneously, to generate any complex patterned acoustic holographic lattice array, such as circular, triangular, square, and liver-shaped complex acoustic holographic lattice patterns.

Subsequently, they used the acoustic radiation force of the acoustic potential well to assemble cells into specific lattice patterns, simulating the anatomical structure of the liver.

In addition, the researcher used expandable primary mouse liver cells in vitro for the first time to deeply study the impact of HAT on the biological effects of primary liver cells. The results showed that primary liver cells under the holographic lattice-based acoustic tweezers produced a large number of self-assembled liver cell spheroids, and compared with traditional 2D and 3D culture models, the diameter of liver cell spheroids significantly increased, and the functions of liver cell protein synthesis metabolism, sugar metabolism, detoxification, etc., were also significantly enhanced.

This work established the foundation for the development of an ex vivo model through AHL techniques, carrying significant implications for future 3D tissue engineering applications.