Institut Necker Enfants Malades, Faculté de Médecine Necker, INSERM U1151, Université de Paris, 160 rue de Vaugirard, 75015 Paris, France.
Medical Genetics Unit, Istituto Giannina Gaslini, 16147 Genova, Italy.
STAR Protoc. 2022 May 27;3(2):101419. doi: 10.1016/j.xpro.2022.101419. eCollection 2022 Jun 17.
Here, we present a standardized protocol for isolation, maintenance, and polarization of the respiratory epithelial primary cells from patient samples acquired from nasal brushing, polyp specimens, or lung explants. This protocol generates a clearly defined polarized layer of epithelial cells on filters, with a good number of ciliated cells and a thin layer of mucus. We detail the steps for samples prepared from patients with cystic fibrosis as well as from subjects without cystic fibrosis.
在这里,我们提供了一种从鼻腔刷取物、息肉标本或肺组织切片等患者样本中分离、维持和极化呼吸道上皮原代细胞的标准化方案。该方案可在过滤器上生成明确界定的极化上皮细胞层,具有大量纤毛细胞和薄的黏液层。我们详细介绍了从囊性纤维化患者和非囊性纤维化患者中制备样本的步骤。