100mm ultra low attachment culture dishes

Multicellular cancer spheroids (MCS) were generated from a heterogeneous parental population by seeding cells growing during log-phase in 100mm ultra-low attachment culture dishes (Corning Inc. New York, NY, USA), culturing them in DMEM /F12 supplemented with 5% FBS then incubated at 37 °C 5% CO₂ incubator. Cells were grown in ultra-low attachment dishes for 2 weeks to make 3D anchorage-independent models. The 3D anchorage-independent models were imaged by Axio Imager 2 Research Microscope (Zeiss, Jena, Germany) and the average size of the spheroids was measured using the Zeiss ZEN Microscope Software for Microscopic Components.

MCF7 cells and their derivatives were seeded into 100-mm ultra-low attachment culture dishes (Corning) in Mammary Epithelial Cell Growth Medium (Lonza) at a density of at 2 × 10⁴ cells per ml.

A canine iPS cell line was generated as previously described (26 (link)) from skin biopsy fibroblast derived from a 6-year old male standard poodle To differentiate iPS cells into Neurospheres (neural progenitor cells), colonies were digested to single cell suspension and re-suspended in media containing DMEM/F12 (27 (link)) (Life Technologies Corp. Grand Island NY), Antibiotic-Antimycotic, B27 (Thermo Fisher Scientific Waltham, MA), 20 ng/mL Recombinant Human FGF-basic (Fibroblast Growth Factor-basic, bFGF), 20 ng/mL Recombinant Human EGF (Epidermal Growth Factor) (PeproTech, Rock Hill, NJ), and 2 μg/mL Heparin (Sigma-Aldrich, St. Louis MO). Dissociated cells were plated in 100 mm ultra-low attachment culture dishes (Corning Inc. Corning, NY) at a density of 4 × 105 cells/mL, on day 10 neurospheres were collected and used for injection or for further differentiation.