Derivation and culture of canine embryonic stem cells

The derivation of canine embryonic stem cells (cESCs) represents a significant achievement and opens the door to further stem cell research and therapies in the dog. Canines share a common environment with humans and exhibit a host of genetic diseases, many of which have human parallels. Thus, the canine model presents unique advantages over other currently used organisms to help develop stem cell therapies in humans. To reveal the therapeutic potential of cESCs further basic research on the molecular mechanisms controlling their pluripotency and self-renewal characteristics is needed. Herein, we present the methods for derivation and culture of cESCs. Following collection of the canine blastocyst, two derivation methods are presented; immunodissection and whole blastocyst explant. These two methods lead to cESCs differing in morphology and subculture techniques. Additional protocols for subculture of established lines, feeder-free culture, and cryopreservation protocols are also described.