Mouse embryonic stem cell ES-E14TG2a from ATCC (CRL-1821) were cultured on 0.1% gelatin-coated plates in DMEM medium containing 15% Fetal Bovine Serum, 1% Penicillin/Streptomycin, 1% Glutamax, 0.1 mM 2-mercaptoethanol, 1% MEM Non-Essential Amino Acids, 1% Sodium Pyruvate, and 1,000 U/ml recombinant leukemia inhibitory factor (LIF).
Wnt3a coated beads were prepared as described88 (link). Localized Wnt3a beads with mESCs were engineered in the 2 cm plates following the previous method15 (link). To achieve one cell and one bead contact, mESCs were seeded at a low concentration before imaging or collecting. After seeding the cells and Wnt3a beads, samples were collected at around 12 h.
Wnt3a coated beads were prepared as described88 (link). Localized Wnt3a beads with mESCs were engineered in the 2 cm plates following the previous method15 (link). To achieve one cell and one bead contact, mESCs were seeded at a low concentration before imaging or collecting. After seeding the cells and Wnt3a beads, samples were collected at around 12 h.
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