Abstract
Direct reprogramming of canine fibroblasts to induced motor neurons in vitro opens the door to molecular characterization, functional analysis, and therapeutic screening of canine age-related neurodegenerative diseases. Here, we present a protocol for generating canine induced motor neurons directly from primary dermal fibroblasts. We provide a step-by-step guide for isolating and maintaining primary dermal fibroblasts from dogs, obtaining a stock of reprogrammable cells, and performing the direct reprogramming process. We then detail procedures for validating the motor neuron identity.