Abstract
BackgroundInduced pluripotent stem cells (iPSCs) have enormous potential in developmental biology studies and in cellular therapies. Although extensively studied and characterized in human and murine models, iPSCs from animals other than mice lack reproducible results.MethodsHerein, we describe the generation of robust iPSCs from equine and bovine cells through lentiviral transduction of murine or human transcription factors Oct4, Sox2, Klf4, and c-Myc and from human and murine cells using similar protocols, even when different supplementations were used. The iPSCs were analyzed regarding morphology, gene and protein expression of pluripotency factors, alkaline phosphatase detection, and spontaneous and induced differentiation.ResultsAlthough embryonic-derived stem cells are yet not well characterized in domestic animals, generation of iPS cells from these species is possible through similar protocols used for mouse or human cells, enabling the use of pluripotent cells from large animals for basic or applied purposes. Herein, we also infer that bovine iPS (biPSCs) exhibit similarity to mouse iPSCs (miPSCs), whereas equine iPSs (eiPSCs) to human (hiPSCs).ConclusionsThe generation of reproducible protocols in different animal species will provide an informative tool for producing in vitro autologous pluripotent cells from domestic animals. These cells will create new opportunities in animal breeding through transgenic technology and will support a new era of translational medicine with large animal models.
Highlights
Induced pluripotent stem cells have enormous potential in developmental biology studies and in cellular therapies
Induced pluripotent stem cells have enormous potential in cellular therapy, reprogramming, and early development approaches as they can differentiate into numerous autologous cell lineages, including the three
Equine adult fibroblasts and adipose tissue mesenchymal cells were transduced with murine or human OSKM, which resulted in induced colonies approximately 3 days after transduction only with human OSKM (Fig. 1) that were independent of Human leukemia inhibitor factor (hLIF)/2i supplementation (Fig. 2)
Summary
Induced pluripotent stem cells (iPSCs) have enormous potential in developmental biology studies and in cellular therapies. Rabbits are considered adequate models for translational research [22, 23]; its in vitro cell culture and pluripotency acquisition still lack further robust characterization [24] In this context, domestic animal species such as swine, small ruminants, and even companion animals such as dogs may provide an important contribution to regenerative and translational medicine. Large farm animals are important models for pre-clinical stem cell therapies due to their physiological and morphological similarity to humans [28, 29], but they may greatly benefit from pluripotency in vitro to the generation of genetically superior or modified organisms for agricultural and biomedical applications, for example, for reproductive sciences [30]
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