Journal: Stem Cell Research & Therapy

Article Title: Transcriptomic insights and feeder-free culturing of porcine expanded potential stem cells from cloned embryos

doi: 10.1186/s13287-025-04627-5

Figure Lengend Snippet: Feeder-free culture condition enhancement for the pEPSC NT line through media components and ECM testing optimization. A Representative morphologies and AP staining images of feeder-free pEPSCs NT (FF-pEPSCs NT ) cultured under different media conditions. XWCLAV: XAV939, WH-4-023, CHIR99021, LIF, Activin A, and vitamin C. F12: DMEM/F12. Scale bars = 200 μm. B Immunofluorescence images of POU5F1 and NANOG protein expression in FF-pEPSCs NT cultured under different culture media conditions. Scale bars = 100 μm. C Expression of core pluripotent genes ( POU5F1 , SOX2 , and NANOG ) in FF-pEPSCs NT cultured under different media conditions. Data represent the mean ± SEM. The relative expression was normalized to that of GAPDH . **** P < 0.0001. n = 3 independent experiments. D Representative morphologies and AP staining images of FF-pEPSCs NT cultured under different ECM conditions 72 h after seeding. Scale bars = 200 μm. E The impacts of various matrix proteins on FF-pEPSCs NT . The percentages of adherent cells at 1.5 h, survival of dissociated cells at 24 h, and proliferation of cells at 72 h after seeding were evaluated. The index represents the number of cells at a specific time point divided by the initial number of seeded cells. n = 3 independent experiments. ** P < 0.01, *** P < 0.001, and **** P < 0.0001. F Immunofluorescence images of POU5F1 and SOX2 protein expression in FF-pEPSCs NT cultured under 1× or 2× Matrigel-coated conditions. Scale bars = 100 μm. G Expression of core pluripotent genes ( POU5F1 , SOX2 , NANOG , and KLF4 ) in FF-pEPSCs NT cultured under various ECM protein-coated conditions. Data represent the mean ± SEM. The relative expression was normalized to that of GAPDH . n = 3 independent experiments. AP alkaline phosphatase, ECM extracellular matrix; pEPSC porcine expanded potential stem cell, VTN-N vitronectin

Article Snippet: The modified pEPSCM was prepared by incorporation using basal medium in a 1:1 mixture of mTeSR TM 1 (100–0276; STEMCELL Technologies, Vancouver, BC, Canada) and DMEM/F-12 (Gibco) supplemented with 1× N2 supplement (Thermo Fisher Scientific), 1× B27 supplement (Thermo Fisher Scientific), 1× NEAA, 1× GlutaMAX supplement (Gibco), 1 × β-mercaptoethanol, 0.5 μM CHIR99021 (S1263; Selleck Chemicals, Houston, TX, USA), 0.3 μM WH-4-023 (S7565; Selleck Chemicals), 2.5 μM XAV939 (X3004), 65.0 μg/mL vitamin C (50-81-7), 10.0 ng/mL leukemia inhibitory factor (LIF) (250-02; PeproTech, Cranbury, NJ, USA), 20.0 ng/mL Activin A (78001; STEMCELL Technologies), 0.3% FBS, and 0.5× Anti-anti (Gibco).

Techniques: Staining, Cell Culture, Immunofluorescence, Expressing

Journal: Nature biomedical engineering

Article Title: Generation of neural organoids for spinal-cord regeneration via the direct reprogramming of human astrocytes

doi: 10.1038/s41551-022-00963-6

Figure Lengend Snippet: a , Schematic for the direct reprogramming of human astrocytes into neurons. b , Schematic diagram of primary screening protocol to convert human astrocytes into neurons using each of 15 small molecules. AM, Astrocyte Medium; NB, Neurobasal medium. c , Representative images of MAP2 + cells treated with each of 15 small molecules and DMSO as control at day 21. Scale bar, 100 μm. d , Quantification of the percentage of induced MAP2 + neurons treated with 15 small molecules (CHIR99021, Y27632, SB431542, XAV939 and SAG, n = 4 independent experiments; the rest, n = 3 independent experiments) and DMSO (n = 4 independent experiments). Data are presented as mean ± SEM.

Article Snippet: From day 3 to day 8, small molecules including CHIR99021 (1.5 μM, Tocris), VPA (5 mM, Cayman), RepSox (1 μM, Tocris), Forskolin (10 μM, Cayman), JQ1 (50 nM, Sigma), ISX-9 (10 μM, Tocris), LDN193189 (100 nM, Stemolecule), TTNPB (0.5 μM, Tocris), Y27632 (10 μM, Tocris), DAPT (2.5 μM, Sigma), SAG (100 nM, EMD Millipore), SB431542 (10 μM, MCE), PD0325901 (10 μM, Tocris), SU5402 (10 μM, Biovision), and XAV939 (10 μM, Tocris) were added to induce neurons cultured with Neural Medium for primary screening.

Techniques: Control