Chang Kung-Chi, Silvestri Francesca, Oliphant Michael U J, Martinez-Gakidis M Angie, Orgill Dennis P, Garber Judy E, Dillon Deborah D, Brugge Joan S
Department of Cell Biology and Ludwig Center at Harvard, Harvard Medical School, 240 Longwood Ave, Boston, MA, 02115, USA.
Department of Surgery, Division of Plastic and Reconstructive Surgery, Brigham & Women's Hospital, 75 Francis St, Boston, MA, 02115, USA.
NPJ Breast Cancer. 2024 Dec 19;10(1):107. doi: 10.1038/s41523-024-00714-7.
Organoid cultures offer a powerful technology to investigate many different aspects of development, physiology, and pathology of diverse tissues. Unlike standard tissue culture of primary breast epithelial cells, breast organoids preserve the epithelial lineages and architecture of the normal tissue. However, existing organoid culture methods are tedious, difficult to scale, and do not robustly retain estrogen receptor (ER) expression and responsiveness in long-term culture. Here, we describe a modified culture method to generate and maintain organoids as suspension cultures in reconstituted basement membrane (™Matrigel). This method improves organoid growth and uniformity compared to the conventional Matrigel dome embedding method, while maintaining the fidelity of the three major epithelial lineages. Using this adopted method, we are able to culture and passage purified hormone sensing (HS) cells that retain ER responsiveness upon estrogen stimulation in long-term culture. This culture system presents a valuable platform to study the events involved in initiation and evolution of ER-positive breast cancer.
类器官培养提供了一项强大的技术，可用于研究多种组织在发育、生理学和病理学方面的许多不同层面。与原发性乳腺上皮细胞的标准组织培养不同，乳腺类器官保留了正常组织的上皮谱系和结构。然而，现有的类器官培养方法繁琐、难以规模化，并且在长期培养中不能稳定地保持雌激素受体（ER）的表达和反应性。在此，我们描述了一种改良的培养方法，以在重组基底膜（基质胶）中作为悬浮培养物来生成和维持类器官。与传统的基质胶穹顶包埋法相比，该方法改善了类器官的生长和均一性，同时保持了三大上皮谱系的保真度。使用这种改进方法，我们能够培养并传代纯化的激素感应（HS）细胞，这些细胞在长期培养中对雌激素刺激保持ER反应性。这种培养系统为研究ER阳性乳腺癌的起始和演变所涉及的事件提供了一个有价值的平台。
