比较离体和体外肠囊性纤维化模型以测量 CFTR 依赖性离子通道活性。
Comparison of ex vivo and in vitro intestinal cystic fibrosis models to measure CFTR-dependent ion channel activity.
机构信息
Department of Pediatric Pulmonology, Wilhelmina Children's Hospital, University Medical Center Utrecht, Utrecht, The Netherlands; Regenerative Medicine Center Utrecht, University Medical Centre Utrecht, Utrecht, The Netherlands.
Department of Pediatric Pulmonology, Erasmus Medical Centre, Sophia Children's Hospital, Rotterdam, The Netherlands.
出版信息
J Cyst Fibros. 2018 May;17(3):316-324. doi: 10.1016/j.jcf.2018.02.007. Epub 2018 Mar 13.
BACKGROUND
New functional assays using primary human intestinal adult stem cell cultures can be valuable tools to study epithelial defects in human diseases such as cystic fibrosis.
METHODS
CFTR-mediated ion transport was measured in rectal organoid-derived monolayers grown from subjects with various CFTR mutations and compared to donor-matched intestinal current measurements (ICM) in rectal biopsies and forskolin-induced swelling of rectal organoids.
RESULTS
Rectal organoid-derived monolayers were generated within four days. Ion transport measurements of CFTR function using these monolayers correlated with ICM and organoid swelling (r = 0.73 and 0.79 respectively). Culturing the monolayers under differentiation conditions enhanced the detection of mucus-secreting cells and was accompanied by reduced CFTR function.
CONCLUSIONS
CFTR-dependent intestinal epithelial ion transport properties can be measured in rectal organoid-derived monolayers of subjects and correlate with donor-matched ICM and rectal organoid swelling.
背景
使用原代人肠成体干细胞培养物进行新的功能测定，可能是研究囊性纤维化等人类疾病中上皮缺陷的有用工具。
方法
在从具有各种 CFTR 突变的受试者中衍生的直肠类器官衍生的单层中测量 CFTR 介导的离子转运，并将其与直肠活检中的供体匹配的肠电流测量（ICM）和 forskolin 诱导的直肠类器官肿胀进行比较。
结果
直肠类器官衍生的单层在四天内生成。使用这些单层进行 CFTR 功能的离子转运测量与 ICM 和类器官肿胀相关（分别为 r=0.73 和 0.79）。在分化条件下培养这些单层可增强对分泌粘液细胞的检测，并伴有 CFTR 功能降低。
结论
可在受试者的直肠类器官衍生单层中测量 CFTR 依赖性肠上皮离子转运特性，并与供体匹配的 ICM 和直肠类器官肿胀相关。
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