Protocol for direct reprogramming of canine fibroblasts to induced motor neurons.

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.