Journal: STAR Protocols

Article Title: Functional assays with human patient-derived enteroid monolayers to assess the human gut barrier

doi: 10.1016/j.xpro.2021.100680

Figure Lengend Snippet: Preparation of Human monolayer EDMs media (100 mL) (stored at 4°C for 2–4 days)

Article Snippet: Embed the living stem cells into a basement membrane matrix (Matrigel) dome and cultured using 50% stem cell-enriched WRN conditioned media (WNT 3a, R-spondin, and Noggin) (prepared from L-WRN cells, ATCC CRL-3276 TM ) containing specific components such as Y27632 (ROCK inhibitor) and SB431542 (an inhibitor for TGF-β type I receptor), nicotinamide, A83-01 (an ACTIVIN /NODAL/TGF-β pathway inhibitor), Epidermal growth factor, and SB202190 (a p38 MAPK inhibitor) to support the growth and long-term cultures of 3D organoids ( , , , ).

Techniques: Concentration Assay

Journal: Cell Reports

Article Title: Labeling and Characterization of Human GLP-1-Secreting L-cells in Primary Ileal Organoid Culture

doi: 10.1016/j.celrep.2020.107833

Figure Lengend Snippet:

Article Snippet: Both media contain the following components in Advanced DMEM:F12 (ADF, Invitrogen): 80% Wnt3A conditioned media, 10% RSPO1 conditioned media, 50 ng/ml murine EGF (Invitrogen), 100 ng/ml murine noggin (BMP-4 antagonist, Peprotech), N2 supplement (Invitrogen), B27 supplement (without vitamin A, Invitrogen), 1 mM N-acetyl-L-cysteine (NAC, Sigma-Aldrich), 500 nM A83-01 (Tocris), 10 nM human [Leu 15 ]-gastrin I (Sigma-Aldrich), 2 mM L-glutamine (Sigma-Aldrich), 10 μM Y-27632 (Tocris) and an additional 50 units/ml penicillin (Sigma-Aldrich) and 50 μg/ml streptomycin (Sigma-Aldrich).

Techniques: Recombinant, Electroporation, RNA Sequencing Assay, Expressing, CRISPR, Software