Reprogramming Equus caballus Fibroblast Cells into Induced Pluripotent Stem Cells

Abstract:

Horses (Equus caballus) are prone to sports injuries that can bring the end of competition, if not life. Induced pluripotent stem cells (iPSCs) are the future of regenerative medicine. The iPSCs are created by taking a skin punch and isolating the fibroblast cells. The fibroblasts are transfected with mRNA to transform them into iPSCs, which can then differentiate into any cell type, such as tendon, cardiac, or nerve cells. These can potentially be used to heal equine injuries. The process used here has been completed successfully with human cells. The hope is that the process to create human iPSCs is transferrable to horses. The goal for this year was to transform Equus caballus fibroblast cells into iPSCs.

Fibroblast cells isolated from a skin punch obtained in 2017 were cultured and transfected with mRNA coding for transcription factors that reprogram them into iPSCs. Different conditions were tested, including multiple cell lines, cell number, volume of media, and amount of transfection reagent. The transfection process was repeated every day for over two weeks, with observation of the cells prompting modification of the process if necessary. After that, another week was spent transforming and expanding the cells.

In one cell line, fibroblasts were observed to transform into starter iPSC colonies. However, enough cells could not be gathered to use for verification of the iPSC state.

Further optimization of conditions will be pursued in future research. The results were promising, despite not being able to create an entire colony of iPSCs.

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