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Modeling colorectal tumorigenesis using the organoids derived from conditionally immortalized mouse intestinal crypt cells (ciMICs)

引用

Genes & Diseases 2021年第6期8卷 814-826页

作者： Xiaoxing Wu Zhaoxia Li Hongyu Zhang Fang He Min Qiao Huaxiu Luo Jing Zhang Meng Zhang Yukun Mao William Wagstaff Yongtao Zhang Changchun Niu Xia Zhao Hao Wang Linjuan Huang Deyao Shi Qing Liu Na Ni Kai Fu Rex C.Haydon Russell R.Reid Hue H.Luu Tong-Chuan He Ziwei Wang Houjie Liang Bing-Qiang Zhang Ning Wang Departments of Gastrointestinal Surgery Medicine/Gastroenterologyand Obstetrics and GynecologyThe First Affiliated Hospital of Chongqing Medical UniversityChongqing400016PR China Molecular Oncology Laboratory Department of Orthopaedic Surgery and Rehabilitation MedicineThe University of Chicago Medical CenterChicagoIL 606037USA Department of Oncology The PLA Rocket Force Characteristic Medical CenterBeijing100088PR China Departments of Burn&Plastic Surgery West China Hospital of Sichuan UniversityChengduSichuan Province610041PR China Department of Orthopaedic Surgery The First Affiliated Hospital of Guangzhou University of Chinese MedicineGuangzhouGuangdong Province510405PR China Departments of Orthopaedic Surgery The Affiliated Zhongnan Hospital of Wuhan UniversityWuhanHubei Province430071PR China Department of Orthopaedic Surgery The Affiliated Hospital of Qingdao UniversityQingdaoShandong Province266000PR China Department of Clinical Laboratory Medicine Chongqing General Hospital Affiliated with the University of Chinese Academy of SciencesChongqing400013PR China ministry of Education Key Laboratory of Diagnostic Medicine and the School of Laboratory Diagnostic MedicineChongqing Medical UniversityChongqing400016PR China Department of Orthopaedics Union Hospital of Tongji Medical CollegeHuazhong University of Science and TechnologyWuhanHubei Province430022PR China Department of Spine Surgery Second Xiangya HospitalCentral South UniversityChangshaHunan Province410008PR China Department of Oncology and Southwest Cancer Center Southwest HospitalArmy Medical UniversityChongqing400038PR China

Intestinal cancers are developed from intestinal epithelial stem cells(ISCs)in intestinal crypts through a multi-step process involved in genetic mutations of oncogenes and tumor suppressor *** play a key role in maintaining the homeostasis of gut *** 2009,Sato et al established a three-dimensional culture system,which mimicked the niche microenvironment by employing the niche factors,and successfully grew crypt ISCs into organoids or mini-guts in *** then,the intestinal organoid technology has been used to delineate cellular signaling in ISC ***,the cultured organoids consist of heterogeneous cell populations,and it was technically challenging to introduce genomic changes into three-dimensional ***,there was a technical necessity to develop a twodimensional ISC culture system for effective genomic *** this study,we established a conditionally immortalized mouse intestinal crypt(ciMIC)cell line by using a piggyBac transposon-based SV40 T antigen expression *** showed that the ciMICs maintained long-term proliferative activity under two-dimensional niche factor-containing culture condition,retained the biological characteristics of intestinal epithelial stem cells,and could form intestinal organoids in three-dimensional *** in vivo cell implantation tests indicated that the ciMICs were non-tumorigenic,the ciMICs overexpressing oncogenic b-catenin and/or KRAS exhibited high proliferative activity and developed intestinal adenoma-like pathological features in ***,these findings strongly suggested that the engineered ciMICs should be used as a valuable tool cell line to dissect the genetic and/or epigenetic underpinnings of intestinal tumorigenesis.

关键词： Cancer modeling Conditional immortalization mini-gut organoids Mouse intestinal crypt(MIC)cells Tumorigenesis

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