The Establishment And Usability Evaluation Of Liver Cancer Organoids And Its Application In Precision Medicine

Objectives:To establish a liver cancer organoids culture platform,supplement the blank of liver cancer organoids research in China,try to improve the success rate of culture by optimizing the culture scheme,and then select liver cancer organoids with typical morphology and stable passage,carry out histological and genetic tests,analyze and explore the feasibility and application of liver cancer organoids.Methods:1.Liver space-occupying patients who diagnosed in the Department of Hepatobiliary Surgery of Zhongshan Hospital Affiliated to Xiamen University from Aug,2018 to Nov,2019 were included in this study.Samples were obtained surgically and cultured in vitro in matrigel-based culture medium.Screened Among them,,and Observed and recorded with a microscope which have the representative growth condition,the typical organoid morphology and the ability of organoid can be successfully passaged 3 times or more.2.Liver cancer organoids with typical morphology under partial microscopy and stable passage were selected,together with their corresponding sample tissues,to make paraffin sections,and Immunohistochemistry(IHC)staining were performed for comparative evaluation and analysis at the histological level.3.Select some hepatocarcinoid organoids with typical morphology and stable passage,together with their corresponding sample tissues and blood,for Whole Exome Sequencing(WES)and Whole Genome Sequencing(WGS)Comparative analysis at the genetic level.Results:A total of 94 liver samples were collected in this experiment,of which 78 were pathologically confirmed as primary liver cancer,6 were metastatic cancer,and 9 were benign liver disease.56 cases of primary liver cancer organoid culture were successful,with a success rate of 71.8%;4 cases of metastatic cancer organoid cultures were successful,the success rate was 66.7%;4 cases of benign liver disease organoid cultures were successful,the success rate was 66.7%.Similar microscopic cell characteristics were observed between organoids and the original specimen.The positive rates of immunohistochemical indicators were close to each other,and the consistent rates of gene expression profiles and mutations were high.Conclusion:A preliminary in vitro organ culture platform for hepatocellular carcinoma was established,and the current organoids culture process and scheme for hepatocellular carcinoma were summarized and optimized.The success rate of culture was higher than that reported in foreign literatures.To get a multi-level assessment for successful liver cancer organoids:it has typical characteristics of liver cancer in histological level,its consistent with the original tumor samples have higher mutation rate in genetic level,two levels of assessment analysis results that the organoids has most of the original tumor tissue properties and characteristics.The results of drug susceptibility screening of liver cancer organoids can provide personalized treatment,but more evidence of clinical efficacy is needed.