The establishment of novel hair follicle development and hair loss model by inducing iPS cells differentiation and aggregation to form a 3D organoid

通过诱导iPS细胞分化和聚集形成3D类器官，建立新型毛囊发育和脱发模型

• 批准号: 21K16203
• 负责人: 周 思奇
• 金额: $ 2.5万
• 依托单位: University of Tsukuba
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Early-Career Scientists
• 财政年份: 2021
• 资助国家: 日本
• 起止时间: 2021-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-21K16203/
• 关键词: iPS cell; 3D organoid; hair follicles; Hair development; organoid; hair loss

In this project, we aim to investigate the hair growth and loss by inducing iPS cell differentiation to form a 3D hair follicle tissue model in vitro.In the first year, we successfully established the iPS culture system in our lab and maintained the ability of pluripotent differentiation after 10 passages of culture. From the second year, we firstly induced iPS cells differentiation into specific cells associated with hair follicles: keratinocytes, fibroblasts, and dermal papilla. Next, we prepared U-shaped 96-well dishes to induce iPS cells forming spheroids. Unfortunately, contrary to our expectations, we found that iPS cells didn't spontaneously aggregate together to form spheroids like other cells.Therefore, we decided to use Matrigel Matrix, an extracellular matrix that has been used in many studies to assist in cell formation of organoids. After mixing iPS cells with Matrigel, the cells were randomly immobilized. After 3 days with culture medium, the formation of organoids was evident, and a significant enlargement was observed at day 5. Morphological changes in cell differentiation were also observed after the addition of growth factors. These results suggest that Matrigel Matrix can provide a suitable 3D environment to help cells form structures similar to real tissues and can be used for cell differentiation.Unfortunately the investigator is unable to complete the further experiments due to a position change. While, these results still provide valuable information and novel method to study the mechanisms of histogenesis and cell differentiation in the future.

本项目旨在通过诱导iPS细胞分化形成三维毛囊组织模型，研究毛发生长和脱落的情况，第一年我们在实验室成功建立了iPS细胞培养体系，培养10代后仍保持多能分化能力。从第二年开始，我们首先诱导iPS细胞分化为与毛囊相关的特定细胞：角质形成细胞、成纤维细胞和毛乳头。接下来，我们制备U形96孔培养皿以诱导iPS细胞形成球状体。不幸的是，与我们的预期相反，我们发现iPS细胞并不像其他细胞那样自发聚集在一起形成球状体。因此，我们决定使用Matrigel Matrix，这是一种细胞外基质，已在许多研究中用于帮助类器官的细胞形成。将iPS细胞与基质胶混合后，随机固定细胞。在用培养基培养3天后，类器官的形成是明显的，并且在第5天观察到显著增大。加入生长因子后，还观察到细胞分化的形态学变化。这些结果表明，Matrigel Matrix可以提供一个合适的三维环境，帮助细胞形成类似于真实的组织的结构，并可用于细胞分化。不幸的是，由于位置的变化，研究者无法完成进一步的实验。这些结果为今后研究组织发生和细胞分化的机制提供了有价值的信息和新的方法。