February 15, 2019
04 pGEMâT Easy vector syst. I Promega A3600 Nanodrop Lite Thermo Fisher TruSeq Nano kit Illumina 20015965 Illumina HiSeqX Illumina DNeasy blood & tissue kit Qiagen 69506 D300e Digital Dispenser Tecan Micromanipulator Narishige Mâ152 Microinjector Narishige IMâ5B Stereomicroscope Leica MZ75 FlexMAP3D BioâRad Laboratories Methods and Prot;ted by pipetting before being resuspended in 5% BME/AO media (15â20,000 organoids mlâ1) and dispensed into 384âwell microplates (Greiner). Each compound was dispensed at 1â100 ÎźM using a TECAN D300e Digital Dispenser, and after 5 days, cell viability was assayed as described above. Paclitaxel (T7402), methotrexate (A6770), crizotinib (PZ0240), and cisplatin (C2210000) were obtained;04 pGEMâT Easy vector syst. I Promega A3600 Nanodrop Lite Thermo Fisher TruSeq Nano kit Illumina 20015965 Illumina HiSeqX Illumina DNeasy blood & tissue kit Qiagen 69506 D300e Digital Dispenser Tecan Micromanipulator Narishige Mâ152 Microinjector Narishige IMâ5B Stereomicroscope Leica MZ75 FlexMAP3D BioâRad Laboratories Methods and Prot
Organoids are self-organizing 3D structures grown from stem cells that recapitulate essential aspects of organ structure and function. Here, we describe a method to establish long-term-expanding human airway organoids from broncho-alveolar resections or lavage material. The pseudostratified airway organoids consist of basal cells, functional multi-ciliated cells, mucus-producing secretory cells, and CC10-secreting club cells. Airway organoids derived from cystic fibrosis (CF) patients allow assessment of CFTR function in an organoid swelling assay. Organoids established from lung cancer resections and metastasis biopsies retain tumor histopathology as well as cancer gene mutations and are amenable to drug screening. Respiratory syncytial virus (RSV) infection recapitulates central disease features, dramatically increases organoid cell motility via the non-structural viral NS2 protein, and preferentially recruits neutrophils upon co-culturing. We conclude that human airway organoids represent versatile models for the in vitro study of hereditary, malignant, and infectious pulmonary disease. Š 2019 The Authors. Published under the terms of the CC BY NC ND 4.0 license.
Sachs, N; Papaspyropoulos, A; Zomer-van Ommen, DD; Heo, I; BĂśttinger, L; Klay, D; Weeber, F; Huelsz-Prince, G; Iakobachvili, N; Amatngalim, GD; de Ligt, J; van Hoeck, A; Proost, N; Viveen, MC; Lyubimova, A; Teeven, L; Derakhshan, S; Korving, J; Begthel, H; Dekkers, JF; Kumawat, K; Ramos, E; van Oosterhout, MF; Offerhaus, GJ; Wiener, DJ; Olimpio, EP; Dijkstra, KK; Smit, EF; van der Linden, M; Jaksani, S; van de Ven, M; Jonkers, J; Rios, AC; Voest, EE; van Moorsel, CH; van der Ent, CK; Cuppen, E; van Oudenaarden, A; Coenjaerts, FE; Meyaard, L; Bont, LJ; Peters, PJ; Tans, SJ; van Zon, JS; Boj, SF; Vries, RG; Beekman, JM; Clevers, H;
Journal: EMBO J.