Isolation and identification of patient-derived liver cancer stem cells and development of personalized treatment strategies

Abstract Background Liver cancer stem cells (LCSCs) are thought to drive the metastasis and recurrence, however, the heterogeneity of molecular markers of LCSCs has hindered the development of effective methods to isolate them. Methods This study introduced an effective approach to isolate and cultu...

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Main Authors: Tingting Guo, Shuai Zhang, Weiping Zeng, Yan Liang, Jinghe Xie, ShouPei Liu, Yaqi Qiu, Yingjie Fu, Yimeng Ou, Keqiang Ma, Bailin Wang, Weili Gu, Yuyou Duan
Format: Article
Language:English
Published: BMC 2024-11-01
Series:Journal of Translational Medicine
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Online Access:https://doi.org/10.1186/s12967-024-05870-9
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author Tingting Guo
Shuai Zhang
Weiping Zeng
Yan Liang
Jinghe Xie
ShouPei Liu
Yaqi Qiu
Yingjie Fu
Yimeng Ou
Keqiang Ma
Bailin Wang
Weili Gu
Yuyou Duan
author_facet Tingting Guo
Shuai Zhang
Weiping Zeng
Yan Liang
Jinghe Xie
ShouPei Liu
Yaqi Qiu
Yingjie Fu
Yimeng Ou
Keqiang Ma
Bailin Wang
Weili Gu
Yuyou Duan
author_sort Tingting Guo
collection DOAJ
description Abstract Background Liver cancer stem cells (LCSCs) are thought to drive the metastasis and recurrence, however, the heterogeneity of molecular markers of LCSCs has hindered the development of effective methods to isolate them. Methods This study introduced an effective approach to isolate and culture LCSCs from human primary liver cancer (HPLC), leveraging mouse embryonic fibroblasts (MEFs) as feeder cells in conjunction with using defined medium. Isolated LCSCs were further characterized by multiple approaches. Transcriptome sequencing data analysis was conducted to identify highly expressed genes in LCSCs and classify different subtypes of liver cancers. Results Total sixteen cell strains were directly isolated from 24 tissues of three types of HPLC without sorting, seven of which could be maintained long-term culture as colony growth on MEFs, which is unique characteristics of stem cells. Even 10 of cloned cells formed the tumors in immunodeficient mice, indicating that those cloned cells were tumorgenic. The histologies and gene expression pattern of human xenografts were very similar to those of HPLC where these cloned cells were isolated. Moreover, putative markers of LCSCs were further verified to all express in cloned cells, confirming that these cells were LCSCs. These cloned LCSCs could be cryopreserved, and still maintained the feature of colony growth on MEFs after the recovery. Compared to suspension culture as conventional approach to culture LCSCs, our approach much better maintained stemness of LCSCs for a long time. To date, these cloned cells could be cultured on MEFs over 12 passages. Moreover, bioinformatics analysis of sequencing data revealed the gene expression profiles in LCSCs, and liver cancers were classified into two subtypes C1 and C2 based on genes associated with the prognosis of LCSCs. Patients of the C2 subtype, which is closely related to the extracellular matrix, were found to be sensitive to treatments such as Cisplatin, Axitinib, JAK1 inhibitors, WNT-c59, Sorafenib, and RO-3306. Conclusion In summary, this effective approach offers new insights into the molecular landscape of human liver cancers, and the identification of the C2 subtype and its unique response to the treatment pave the way for the creation of more effective, personalized therapeutic strategies.
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spelling doaj-art-1dd9e555d1f544f3902e65ff9c35ff0d2025-08-20T02:33:06ZengBMCJournal of Translational Medicine1479-58762024-11-0122112110.1186/s12967-024-05870-9Isolation and identification of patient-derived liver cancer stem cells and development of personalized treatment strategiesTingting Guo0Shuai Zhang1Weiping Zeng2Yan Liang3Jinghe Xie4ShouPei Liu5Yaqi Qiu6Yingjie Fu7Yimeng Ou8Keqiang Ma9Bailin Wang10Weili Gu11Yuyou Duan12Laboratory of Stem Cells and Translational Medicine, Institute for Medical Research, The Second Affiliated Hospital, School of Medicine, South China University of TechnologyDepartment of Gastroenterology and Hepatology, Guangzhou Digestive Disease Center, Guangzhou First People’s HospitalLaboratory of Stem cells and Translational Medicine, Institutes for Life Sciences, School of Medicine, South China University of TechnologyLaboratory of Stem cells and Translational Medicine, Institutes for Life Sciences, School of Medicine, South China University of TechnologySchool of Biomedical Sciences and Engineering, Guangzhou International Campus, South China University of TechnologyLaboratory of Stem Cells and Translational Medicine, Institute for Medical Research, The Second Affiliated Hospital, School of Medicine, South China University of TechnologyLaboratory of Stem cells and Translational Medicine, Institutes for Life Sciences, School of Medicine, South China University of TechnologyLaboratory of Stem Cells and Translational Medicine, Institute for Medical Research, The Second Affiliated Hospital, School of Medicine, South China University of TechnologyDepartment of General Surgery, The First Affiliated Hospital of Guangdong Pharmaceutical UniversityDepartment of Hepatobiliary Pancreatic Surgery, Huadu District People’s Hospital of GuangzhouDepartment of General Surgery, Guangzhou Red Cross Hospital, Jinan UniversityDepartment of Gastroenterology and Hepatology, Guangzhou Digestive Disease Center, Guangzhou First People’s HospitalLaboratory of Stem Cells and Translational Medicine, Institute for Medical Research, The Second Affiliated Hospital, School of Medicine, South China University of TechnologyAbstract Background Liver cancer stem cells (LCSCs) are thought to drive the metastasis and recurrence, however, the heterogeneity of molecular markers of LCSCs has hindered the development of effective methods to isolate them. Methods This study introduced an effective approach to isolate and culture LCSCs from human primary liver cancer (HPLC), leveraging mouse embryonic fibroblasts (MEFs) as feeder cells in conjunction with using defined medium. Isolated LCSCs were further characterized by multiple approaches. Transcriptome sequencing data analysis was conducted to identify highly expressed genes in LCSCs and classify different subtypes of liver cancers. Results Total sixteen cell strains were directly isolated from 24 tissues of three types of HPLC without sorting, seven of which could be maintained long-term culture as colony growth on MEFs, which is unique characteristics of stem cells. Even 10 of cloned cells formed the tumors in immunodeficient mice, indicating that those cloned cells were tumorgenic. The histologies and gene expression pattern of human xenografts were very similar to those of HPLC where these cloned cells were isolated. Moreover, putative markers of LCSCs were further verified to all express in cloned cells, confirming that these cells were LCSCs. These cloned LCSCs could be cryopreserved, and still maintained the feature of colony growth on MEFs after the recovery. Compared to suspension culture as conventional approach to culture LCSCs, our approach much better maintained stemness of LCSCs for a long time. To date, these cloned cells could be cultured on MEFs over 12 passages. Moreover, bioinformatics analysis of sequencing data revealed the gene expression profiles in LCSCs, and liver cancers were classified into two subtypes C1 and C2 based on genes associated with the prognosis of LCSCs. Patients of the C2 subtype, which is closely related to the extracellular matrix, were found to be sensitive to treatments such as Cisplatin, Axitinib, JAK1 inhibitors, WNT-c59, Sorafenib, and RO-3306. Conclusion In summary, this effective approach offers new insights into the molecular landscape of human liver cancers, and the identification of the C2 subtype and its unique response to the treatment pave the way for the creation of more effective, personalized therapeutic strategies.https://doi.org/10.1186/s12967-024-05870-9Liver cancerCancer stem cellsPersonalized therapyIsolation and culture
spellingShingle Tingting Guo
Shuai Zhang
Weiping Zeng
Yan Liang
Jinghe Xie
ShouPei Liu
Yaqi Qiu
Yingjie Fu
Yimeng Ou
Keqiang Ma
Bailin Wang
Weili Gu
Yuyou Duan
Isolation and identification of patient-derived liver cancer stem cells and development of personalized treatment strategies
Journal of Translational Medicine
Liver cancer
Cancer stem cells
Personalized therapy
Isolation and culture
title Isolation and identification of patient-derived liver cancer stem cells and development of personalized treatment strategies
title_full Isolation and identification of patient-derived liver cancer stem cells and development of personalized treatment strategies
title_fullStr Isolation and identification of patient-derived liver cancer stem cells and development of personalized treatment strategies
title_full_unstemmed Isolation and identification of patient-derived liver cancer stem cells and development of personalized treatment strategies
title_short Isolation and identification of patient-derived liver cancer stem cells and development of personalized treatment strategies
title_sort isolation and identification of patient derived liver cancer stem cells and development of personalized treatment strategies
topic Liver cancer
Cancer stem cells
Personalized therapy
Isolation and culture
url https://doi.org/10.1186/s12967-024-05870-9
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