Journal: iScience

Article Title: Neuronal reprogramming of mouse and human fibroblasts using transcription factors involved in suprachiasmatic nucleus development

doi: 10.1016/j.isci.2024.109051

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Article Snippet: To generate iNs, cells were cultured for 2 days in tetracycline-free FBS-containing medium with the addition of doxycycline (1 μg/mL, Sigma-Aldrich) before changing to optimized induced neuronal conversion medium, containing a 50:50 base of DMEM:F12/Neurobasal containing the following supplements: N2 supplement, B27 supplement (both 1 ×; Gibco, Thermo Fisher Scientific), doxycycline (2 μg/mL, Sigma-Aldrich), Laminin (1 μg/mL, Life Technologies, Carlsbad, CA, USA), dibutyryl cyclic-AMP (500 μg/mL, Sigma-Aldrich), human recombinant Noggin (150 ng/mL; Peprotech), LDN-193189 (0.5 μM; Cayman Chemical Co, Ann Arbor, MI, USA), A83-1 (0.5 μM; Stemgent, Lexington, MA, USA), CHIR99021 (3 μM, LC Laboratories, Woburn, MA, USA), Forskolin (5 μM, LC Laboratories) and SB-431542 (10 μM; Cayman Chemical Co).

Techniques: Virus, Recombinant, Fluorescence, Cell Culture, Sample Prep, SYBR Green Assay, Protease Inhibitor, Imaging, Synthesized, Software