Todhunter Michael E, Miyano Masaru, Moolamalla Divya S, Filippov Aleksandr, Sayaman Rosalyn W, LaBarge Mark A
Department of Population Sciences, Beckman Research Institute, City of Hope, 1500 E. Duarte Road, Duarte, CA 91010, USA.
iScience. 2021 Mar 1;24(4):102253. doi: 10.1016/j.isci.2021.102253. eCollection 2021 Apr 23.
A long-standing constraint on organoid culture is the need to add exogenous substances to provide hydrogel matrix, which limits the study of fully human or fully native organoids. This paper introduces an approach to culture reconstituted mammary organoids without the impediment of exogenous matrix. We enclose organoids in nanoliter-scale, topologically enclosed, fluid compartments surrounded by agar. Organoids cultured in these "microcontainers" appear to secrete enough extracellular matrix to yield a self-sufficient microenvironment without exogenous supplements. In microcontainers, mammary organoids exhibit contractility and a high-level, physiological, myoepithelial (MEP) behavior that has not been previously reported in reconstituted organoids. The presence of contractility suggests that microcontainers elicit MEP functional differentiation, an important milestone. Microcontainers yield thousands of substantially identical and individually trackable organoids within a single culture vessel, enabling longitudinal studies and statistically powerful experiments, such as the evaluation of small effect sizes. Microcontainers open new doors for researchers who rely on organoid models.
长期以来，类器官培养的一个限制因素是需要添加外源物质来提供水凝胶基质，这限制了对完全人类或完全天然类器官的研究。本文介绍了一种在没有外源基质阻碍的情况下培养重组乳腺类器官的方法。我们将类器官封装在由琼脂包围的纳升规模、拓扑封闭的流体隔室中。在这些“微容器”中培养的类器官似乎能分泌足够的细胞外基质，从而在无需外源补充的情况下产生一个自给自足的微环境。在微容器中，乳腺类器官表现出收缩性以及高水平的生理性肌上皮（MEP）行为，这在重组类器官中此前尚未有报道。收缩性的存在表明微容器引发了MEP功能分化，这是一个重要的里程碑。微容器在单个培养容器中可产生数千个基本相同且可单独追踪的类器官，从而能够进行纵向研究以及具有强大统计学效力的实验，比如对微小效应大小的评估。微容器为依赖类器官模型的研究人员打开了新的大门。
