Department of Pediatric Pulmonology, Wilhelmina Children's Hospital, University Medical Center Utrecht, Utrecht University, Member of ERN-LUNG, Utrecht EA 3584, the Netherlands; Regenerative Medicine Center Utrecht, University Medical Center Utrecht, Utrecht University, Utrecht CB 3584, the Netherlands.
Department of Pediatric Pulmonology, Wilhelmina Children's Hospital, University Medical Center Utrecht, Utrecht University, Member of ERN-LUNG, Utrecht EA 3584, the Netherlands; Regenerative Medicine Center Utrecht, University Medical Center Utrecht, Utrecht University, Utrecht CB 3584, the Netherlands; Centre for Living Technologies, Alliance TU/e, WUR, UU, UMC Utrecht, Utrecht CB 3584, the Netherlands.
STAR Protoc. 2023 Sep 15;4(3):102337. doi: 10.1016/j.xpro.2023.102337. Epub 2023 Jun 12.
We present a protocol to generate organoids from air-liquid-interface (ALI)-differentiated nasal epithelia. We detail their application as cystic fibrosis (CF) disease model in the cystic fibrosis transmembrane conductance regulator (CFTR)-dependent forskolin-induced swelling (FIS) assay. We describe steps for isolation, expansion and cryostorage of nasal brushing-derived basal progenitor cells, and their differentiation in ALI cultures. Furthermore, we detail the conversion of differentiated epithelial fragments into organoids of healthy controls and CF subjects for validating CFTR function and modulator responses. For complete details on the use and execution of this protocol, please refer to Amatngalim et al..
我们提出了一种从气液界面 (ALI) 分化的鼻上皮生成类器官的方案。我们详细介绍了它们在囊性纤维化跨膜电导调节剂 (CFTR) 依赖性福斯柯林诱导肿胀 (FIS) 测定中作为囊性纤维化 (CF) 疾病模型的应用。我们描述了从鼻刷分离、扩增和低温保存基底祖细胞的步骤，以及它们在 ALI 培养物中的分化。此外，我们详细介绍了将分化的上皮片段转化为健康对照和 CF 患者的类器官的方法，以验证 CFTR 功能和调节剂反应。有关使用和执行此方案的完整详细信息，请参阅 Amatngalim 等人的研究。
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