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

Genes and Diseases

Tong-Chuan He¹  Russell R. Reid²  Houjie Liang²  Deyao Shi³  Yongtao Zhang⁴  Na Ni⁵  Ning Wang⁶  Hao Wang⁷  Jing Zhang⁷  Yukun Mao⁷  Min Qiao⁷  Zhaoxia Li⁷  Changchun Niu⁸  Hongyu Zhang⁹  Xia Zhao^1,10  Meng Zhang^1,10  Xiaoxing Wu^1,10  Huaxiu Luo^1,10  Fang He^1,10  Linjuan Huang^1,11  Ziwei Wang^1,11  Hue H. Luu^1,11  Kai Fu^1,11  Bing-Qiang Zhang^1,11  William Wagstaff^1,11  Qing Liu^1,11  Rex C. Haydon^1,11 

[1] Department of Clinical Laboratory Medicine, Chongqing General Hospital Affiliated with the University of Chinese Academy of Sciences, Chongqing, 400013, PR China;Department of Orthopaedic Surgery, The Affiliated Hospital of Qingdao University, Qingdao, Shandong Province, 266000, PR China;Department of Orthopaedic Surgery, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, Guangdong Province, 510405, PR China;;Departments of Burn &Departments of Orthopaedic Surgery, The Affiliated Zhongnan Hospital of Wuhan University, Wuhan, Hubei Province, 430071, PR China;Ministry of Education Key Laboratory of Diagnostic Medicine, and the School of Laboratory Diagnostic Medicine, Chongqing Medical University, Chongqing, 400016, PR China;Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center, Chicago, IL 606037, USA;Plastic Surgery, West China Hospital of Sichuan University, Chengdu, Sichuan Province, 610041, PR China;Department of Oncology, The PLA Rocket Force Characteristic Medical Center, Beijing, 100088, PR China;Departments of Gastrointestinal Surgery, Medicine/Gastroenterology, and Obstetrics and Gynecology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, PR China;Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center, Chicago, IL 606037, USA;

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

DOI  :  

来源: DOAJ

【 摘 要 】

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 genes. ISCs play a key role in maintaining the homeostasis of gut epithelium. In 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 vitro. Since then, the intestinal organoid technology has been used to delineate cellular signaling in ISC biology. However, the cultured organoids consist of heterogeneous cell populations, and it was technically challenging to introduce genomic changes into three-dimensional organoids. Thus, there was a technical necessity to develop a two-dimensional ISC culture system for effective genomic manipulations. In this study, we established a conditionally immortalized mouse intestinal crypt (ciMIC) cell line by using a piggyBac transposon-based SV40 T antigen expression system. We 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 culture. While in vivo cell implantation tests indicated that the ciMICs were non-tumorigenic, the ciMICs overexpressing oncogenic β-catenin and/or KRAS exhibited high proliferative activity and developed intestinal adenoma-like pathological features in vivo. Collectively, 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.

【 授权许可】

Unknown