Plates Coating With Poly-L-Ornithine and Laminin

Culture plates were pre-covered with poly-L-ornithine (Sigma P3655) and laminin (Thermo Fisher 23017015) for hGSC culture and passage as previously described (Han et al., 2017). In brief, each 100 mm dish was treated with 10 ml 0.5 μg/ml poly-L-ornithine (10 mg/ml stock concentration dissolved in water) and maintained at room temperature for 16 h on a flat, clean tabletop. The following day, 1 × PBS (Hyclone SH30258.01) was used applied to wash the dishes after the poly-L-ornithine supernatant was discarded. Then, 10 ml 1 × PBS containing 5 μg/ml laminin was added to the dish and was left to keep infiltrating for at least 16 h at room temperature. Finally, the coated dishes were stored at −20°C. Before cell culture, the coated dishes were incubated at 37°C, and the supernatant was discarded.