英国剑桥大学Philip H. Jones团队近期取得重要工作进展，他们研究发现，自我维持的长期3D上皮样培养揭示食管上皮克隆扩增的驱动因素。相关研究成果2024年9月23日在线发表于《自然—遗传学》杂志上。

据介绍，衰老的上皮细胞被体细胞突变定植，这些突变受到内在和外在因素的选择影响。缺乏合适的培养体系减缓了对这一过程和其他长期生物过程的研究。

研究人员描述了上皮样细胞，这是一种培养多种小鼠和人类上皮细胞的简单、经济有效的方法。食管上皮样细胞在不传代的情况下自我维持至少1年，保持3D结构，其中增殖的基底细胞分化为基底上细胞，最终脱落并保持基因组稳定性。5个月的实时成像显示，上皮样细胞在体内复制细胞动力学。上皮样细胞支持基因操作，能够研究3D上皮细胞中的突变细胞竞争和选择，并显示抗癌治疗如何调节转化细胞和野生型细胞之间的竞争。

最后，有针对性的CRISPR-Cas9筛选表明，上皮样细胞再现了衰老人类食管中的突变基因选择，并确定了克隆扩增的其他驱动因素，解决了支撑竞争适应性的遗传网络。

附：英文原文

Title: Self-sustaining long-term 3D epithelioid cultures reveal drivers of clonal expansion in esophageal epithelium

Author: Herms, Albert, Fernandez-Antoran, David, Alcolea, Maria P., Kalogeropoulou, Argyro, Banerjee, Ujjwal, Piedrafita, Gabriel, Abby, Emilie, Valverde-Lopez, Jose Antonio, Ferreira, Ins S., Caseda, Irene, Bejar, Maria T., Dentro, Stefan C., Vidal-Notari, Sara, Ong, Swee Hoe, Colom, Bartomeu, Murai, Kasumi, King, Charlotte, Mahbubani, Krishnaa, Saeb-Parsy, Kourosh, Lowe, Alan R., Gerstung, Moritz, Jones, Philip H.

Issue&Volume: 2024-09-23

Abstract: Aging epithelia are colonized by somatic mutations, which are subjected to selection influenced by intrinsic and extrinsic factors. The lack of suitable culture systems has slowed the study of this and other long-term biological processes. Here, we describe epithelioids, a facile, cost-effective method of culturing multiple mouse and human epithelia. Esophageal epithelioids self-maintain without passaging for at least 1year, maintaining a three-dimensional structure with proliferative basal cells that differentiate into suprabasal cells, which eventually shed and retain genomic stability. Live imaging over 5months showed that epithelioids replicate in vivo cell dynamics. Epithelioids support genetic manipulation and enable the study of mutant cell competition and selection in three-dimensional epithelia, and show how anti-cancer treatments modulate competition between transformed and wild-type cells. Finally, a targeted CRISPR–Cas9 screen shows that epithelioids recapitulate mutant gene selection in aging human esophagus and identifies additional drivers of clonal expansion, resolving the genetic networks underpinning competitive fitness.

DOI: 10.1038/s41588-024-01875-8

Source: https://www.nature.com/articles/s41588-024-01875-8