Canine iPSC Neurosphere Differentiation

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.

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Chow L., McGrath S., de Arruda Saldanha C., Whalen L.R., Packer R, & Dow S. (2020). Generation of Neural Progenitor Cells From Canine Induced Pluripotent Stem Cells and Preliminary Safety Test in Dogs With Spontaneous Spinal Cord Injuries. Frontiers in Veterinary Science, 7, 575938.

Publication 2020

DMEM/F12 Recombinant Human FGF-basic Heparin 100 mm ultra-low attachment culture dishes

Antibiotic Biopsy Canine Cell line Cells Dishes Epidermal growth factor Fibroblast Heparin Human Ips cells Life island Male Neural progenitor cells Recombinant human basic fibroblast growth factor Skin

Corresponding Organization : Colorado State University

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Variable analysis

independent variables

Dissociated cells were plated in 100 mm ultra-low attachment culture dishes at a density of 4 × 10^5 cells/mL
Media containing DMEM/F12, Antibiotic-Antimycotic, B27, 20 ng/mL Recombinant Human FGF-basic (Fibroblast Growth Factor-basic, bFGF), 20 ng/mL Recombinant Human EGF (Epidermal Growth Factor), and 2 μg/mL Heparin

dependent variables

Differentiation of iPS cells into Neurospheres (neural progenitor cells)

control variables

Canine iPS cell line generated from skin biopsy fibroblasts derived from a 6-year old male standard poodle

controls

No positive or negative controls explicitly mentioned.

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