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1.
Chinese Journal of Hepatology ; (12): 532-537, 2023.
Article in Chinese | WPRIM | ID: wpr-986164

ABSTRACT

Objective: To explore a simple and feasible method for the isolation and purification of hepatocytes, hepatic stellate cells (HSC), and lymphocytes from mice. Methods: The cell suspension was obtained from male C57bl/6 mice by hepatic perfusion through the portal vein digestion method and then isolated and purified by discontinuous Percoll gradient centrifugation. Trypan blue exclusion was used to determine cell viability. Glycogen staining, cytokeratin 18, and transmission electron microscopy were used to identify hepatic cells. Immunofluorescence was used to detect α-smooth muscle actin combined with desmin in HSCs. Flow cytometry was used to analyze lymphocyte subsets in the liver. Results: After isolation and purification, about 2.7×10(7) hepatocytes, 5.7×10(5) HSCS, and 4.6×106 hepatic mononuclear cells were obtained from the liver of mice with a body weight of about 22g. The cell survival rate in each group was > 95%. Hepatocytes were apparent in glycogen deposited purple-red granules and cytokeratin 18. Electron microscopy showed that there were abundant organelles in hepatocytes and tight junctions between cells. HSC had expressed α-smooth muscle actin and desmin. Flow cytometry showed hepatic mononuclear cells, including lymphocyte subsets such as CD4, CD8, NKs, and NKTs. Conclusion: The hepatic perfusion through the portal vein digestion method can isolate multiple primary cells from the liver of mice at once and has the features of simplicity and efficiency.


Subject(s)
Male , Mice , Animals , Keratin-18 , Actins , Desmin , Liver , Hepatocytes , Hepatic Stellate Cells
2.
China Journal of Chinese Materia Medica ; (24): 2892-2898, 2018.
Article in Chinese | WPRIM | ID: wpr-687369

ABSTRACT

The theory of medicinal properties of traditional Chinese medicine (TCM) is one of the core theories of TCM, which is an important basis for guiding the clinical application of TCM and the key supporting point for the modernization of TCM. How to study the modernization of TCM under the guidance of traditional Chinese medicine theory has become a great challenge for modern researchers. Currently, the researches in TCM mainly focus on chemical studies such as on active ingredients and effective components, lacking the guidance of traditional Chinese medicine theory, and losing the inherent characteristics of TCM. Therefore, the combination of the theory of TCM and the modernization of TCM and the establishment of research mode that is in line with the characteristics of modernization of traditional Chinese medicine under guidance by the theory of medicinal properties of TCM shall be the top priority in modern Chinese medicine research. The property and flavor of TCM are closely related to its chemical compositions, and also have certain regularity. The researches on the chemical components of TCM, systematic design of property and flavor, as well as the separation and combination of TCM property and flavor are beneficial to the overall analysis of the characteristics of TCM, showing great significance to inherit and carry forward TCM. The pharmacological effects about separation and combination methods based on TCM property and flavor were reviewed in this study, providing theoretical reference for the development of new drugs and the application of TCM.

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