Generation of Leukemia Inhibitory Factor and Basic Fibroblast Growth Factor-Dependent Induced Pluripotent Stem Cells from Canine Adult Somatic Cells

Abstract

For more than thirty years, the dog has been used as a model for human diseases. Despite efforts made to develop canine embryonic stem cells, success has been elusive. Here, we report the generation of canine induced pluripotent stem cells (ciPSCs) from canine adult fibroblasts, which we accomplished by introducing human OCT4, SOX2, c-MYC, and KLF4. The ciPSCs expressed critical pluripotency markers and showed evidence of silencing the viral vectors and normal karyotypes. Microsatellite analysis indicated that the ciPSCs showed the same profile as the donor fibroblasts but differed from cells taken from other dogs. Under culture conditions favoring differentiation, the ciPSCs could form cell derivatives from the ectoderm, mesoderm, and endoderm. Further, the ciPSCs required leukemia inhibitory factor and basic fibroblast growth factor to survive, proliferate, and maintain pluripotency. Our results demonstrate an efficient method for deriving canine pluripotent stem cells, providing a powerful platform for the development of new models for regenerative medicine, as well as for the study of the onset, progression, and treatment of human and canine genetic diseases.

Preclinical Derivation and Imaging of Autologously Transplanted Canine Induced Pluripotent Stem Cells

“…More recently, the derivation of ciPSCs from canine embryonic fibroblasts and testicular tissue has been described (25,26). However, the ciPSCs derived in these preliminary studies were not shown capable of teratoma formation nor were compared with cESCs in terms of comprehensive analysis for markers of pluripotency.…”

Defining the Diversity of Phenotypic Respecification Using Multiple Cell Lines and Reprogramming Regimens

“…It was long thought likely that reprogrammed cells arose from cryptic populations of canonical stem cells of varying potency contaminating the main population, but direct reprogramming experiments from many sources to produce induced pluripotent stem cells [6,[33][34][35][36][37][38], neurons [7,[24][25][26][27]39,40], hepatocytes [9,41], cardiomyocytes [8,13,42], and other induced cell types have since largely disproved this proposition.…”

Endothelial Cardiac Cell Therapy

“…iPSCs have been created from adult somatic cells of many small and large animals including mice, 15 rats, 16 monkeys, 17 dogs, 18 pigs, 13,14 and humans, 19 confirming the universality of transgenes for reprogramming. Species such as the pig, whose body and organ size, anatomy, longevity, and physiology resemble the human more than the mouse, are important in transplantation biology.…”