Standardizing the Isolation, Culture‐Expansion, and Cryopreservation of Canine Umbilical‐Cord Derived Mesenchymal Stromal Cells

Abstract

Mesenchymal stromal cells (MSCs) hold great promise in the field of regenerative medicine due to their ability to enable functional recovery in inflammatory injuries such as Crohn's disease and osteoarthritis. Currently, the MSC literature is focused on rodents for preclinical models of disease or clinical testing in human patients. We are interested in using the canine as a large animal model of human diseases. In this regard, the canine MSC literature lags behind human and rodent MSC, and most work on canine MSCs has focused upon bone marrow and adipose tissue‐derived with wide variation in isolation techniques. From surveying the literature and our pilot work, canine MSCs are more difficult to culture than human or rodent MSCs, have longer population doubling times, and earlier senesce. Here, a reproducible protocol for isolation and expansion of canine umbilical cord MSCs is provided. This method produces 10× greater yield per gram of tissue than other published methods. Gelatin‐coating of tissue culture plates greatly increases colony‐forming units‐fibroblast efficiency and decreases population doubling times. In addition, MSCs were able to be cultured longer before senescing, thereby increasing their cumulative population doublings. In summary, our understanding of the culture conditions needed for canine MSCs lags behind that of other species. Gelatin‐coating of tissue culture plates appears to be important for reproducible canine MSC culture. Currently, we are testing differentiation and characterization protocols to offer as standardized methods for the field.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.