Vallone Valeria Fernandez, Leprovots Morgane, Vassart Gilbert, Garcia Marie-Isabelle
Institut de Recherche Interdisciplinaire en Biologie Humaine et Moléculaire (IRIBHM), Faculty of Medicine, Université Libre de Bruxelles ULB, Brussels, Belgium.
Bio Protoc. 2017 Jan 5;7(1):e2088. doi: 10.21769/BioProtoc.2088.
The tri-dimensional culture, initially described by Sato (2009) in order to isolate and characterize epithelial stem cells of the adult small intestine, has been subsequently adapted to many different organs. One of the first examples was the isolation and culture of antral stem cells by Barker (2010), who efficiently generated organoids that recapitulate the mature pyloric epithelium . This approach is suitable and promising to study gastric function in homeostasis as well as in disease. We have adapted Barker's protocol to compare homeostatic and regenerating tissues and here, we meticulously describe, step by step, the isolation and culture of antral glands as well as the isolation of single cells from antral glands that might be useful for culture after cell sorting as an example (Fernandez Vallone , 2016 ).
三维培养最初由佐藤(2009年)描述,用于分离和鉴定成年小肠的上皮干细胞,随后已应用于许多不同器官。最早的例子之一是巴克(2010年)对胃窦干细胞的分离和培养,他高效地生成了模拟成熟幽门上皮的类器官。这种方法适用于研究稳态和疾病状态下的胃功能,且前景广阔。我们采用了巴克的方案来比较稳态组织和再生组织,在此,我们逐步详细描述胃窦腺的分离和培养,以及从胃窦腺中分离单个细胞的方法,这些单个细胞经细胞分选后可能有助于培养(费尔南德斯·瓦洛内,2016年)。