School of Life Science and Technology, ShanghaiTech University, Shanghai 201210, China.
University of Chinese Academy of Sciences, Beijing, China.
STAR Protoc. 2021 Aug 27;2(3):100778. doi: 10.1016/j.xpro.2021.100778. eCollection 2021 Sep 17.
We recently established an culture system in which mammary gland organoid undergoes directional migration in response to an FGF10 concentration gradient. Here, we describe a step-by-step protocol for preparing organoids, the setup of the 3D culture system, and the image acquisition approach. The technical difficulties in conducting the 3D migration assay are choosing epithelial organoids of appropriate sizes and manually paring organoids and beads pre-soaked in FGF10 within a desirable distance (∼100 μm). For complete details on the use and execution of this protocol, please refer to Lu et al. (2020).
我们最近建立了一个培养体系，在该体系中，乳腺类器官会定向迁移以响应 FGF10 浓度梯度。在这里，我们将描述制备类器官的分步方案、3D 培养系统的设置以及图像采集方法。进行 3D 迁移分析的技术难点在于选择大小合适的上皮类器官，以及在理想距离（约 100μm）内手动将类器官和预先浸泡在 FGF10 中的珠子配对。如需详细了解本方案的使用和实施，请参考 Lu 等人（2020 年）。
