Modeling colorectal tumorigenesis using the organoids derived from conditionally immortalized mouse intestinal crypt cells (ciMICs)
| Autor: | Tong-Chuan He, Jing Zhang, Fang He, Rex C. Haydon, Hongyu Zhang, Meng Meng Zhang, Na Ni, Xia Zhao, Yongtao Zhang, Huaxiu Luo, Linjuan Huang, William Wagstaff, Kai Fu, Bing-Qiang Zhang, Changchun Niu, Ning Wang, Hao Hao Wang, Russell R. Reid, Houjie Liang, Deyao Shi, Yukun Mao, Hue H. Luu, Qing Liu, Ziwei Wang, Xiaoxing Wu, Zhaoxia Li, Min Qiao |
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| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Medicine (General) Cell signaling Cell Crypt QH426-470 Biology medicine.disease_cause Cancer modeling digestive system Biochemistry Mini-gut organoids 03 medical and health sciences R5-920 0302 clinical medicine Full Length Article Genetics Organoid medicine Epigenetics Molecular Biology Genetics (clinical) Cell Biology Cell biology Conditional immortalization 030104 developmental biology medicine.anatomical_structure Cell culture 030220 oncology & carcinogenesis Tumorigenesis Stem cell Carcinogenesis Mouse intestinal crypt (MIC) cells |
| Zdroj: | Genes & Diseases Genes and Diseases, Vol 8, Iss 6, Pp 814-826 (2021) |
| ISSN: | 2352-3042 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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