Generation, Characterization, and Multilineage Potency of Mesenchymal-Like Progenitors Derived from Equine Induced Pluripotent Stem Cells

Abstract

Multipotent mesenchymal stromal cells (MSCs) are more and more frequently used to treat orthopedic injuries in horses. However, these cells are limited in their expandability and differentiation capacity. Recently, the first equine-induced pluripotent stem cell (iPSC) lines were reported by us [ 1 ]. In vitro differentiation of iPSCs into MSC-like cells is an attractive alternative to using MSCs derived from other sources, as a much larger quantity of patient-specific cells with broad differentiation potential could be generated. However, the differentiation capacity of iPSCs to MSCs and the potential for use in tissue engineering have yet to be explored. In this study, equine iPSCs were induced to differentiate into an MSC-like population. Upon induction, the iPSCs changed morphology toward spindle-shaped cells similar to MSCs. The ensuing iPSC-MSCs exhibited downregulation of pluripotency-associated genes and an upregulation of MSC-associated genes. In addition, the cells expressed the same surface markers as MSCs derived from equine umbilical cord blood. We then assessed the multilineage differentiation potential of iPSC-MSCs. Although chondrogenesis was not achieved after induction with transforming growth factor-beta 3 (TGFβ3) and/or bone morphogenic protein 4 (BMP-4) in 3D pellet culture, mineralization characteristic of osteogenesis and lipid droplet accumulation characteristic of adipogenesis were observed after chemical induction. We demonstrate a protocol for the derivation of MSC-like progenitor populations from equine iPS cells.

Induced Pluripotent Stem Cells in Dental and Nondental Tissue Regeneration: A Review of an Unexploited Potential

“…Moreover, the osteogenic potential of iPSCs-MSCs derived from either human deciduous teeth or human DF was higher than that of osteoinduced SHED [94]. iPSCs-MSCs derived from equine fibroblast iPSCs were compared to MSCs derived from newborn foals' umbilical cord blood (CB-MSCs) [96]. Von Kossa and alizarin red staining of iPSCs-MSCs showed early mineralization indicating early osteogenesis which was consistent with the results obtained from CB-MSCs.…”

Biocompatible Three-Dimensional Printed Thermoplastic Scaffold for Osteoblast Differentiation of Equine Induced Pluripotent Stem Cells

“…As we were not able to extract live cells from the scaffold, it was not possible to quantify the efficiency of differentiation of the iPSCs, for example using flow cytometry. Other methods using an intermediate differentiation step to mesenchymal stem cells (MSCs) 44 and removal of any undifferentiated iPSCs (using cell sorting) prior to 3D differentiation may help to reduce the safety concerns of using iPSC derived products clinically. Future work to determine the optimal duration of 3D differentiation is also required as in this study we only examined differentiation after 21 days.…”

The Proangiogenic Potential of Mesenchymal Stem Cells and Their Therapeutic Applications

“…This method of cellular reprogramming has been shown to be universal and can be applied to a variety of cell types such as B-cells [16], liver cells [17], and umbilical cord blood mononuclear cells [18]. Moreover, iPS cells have been generated from different species such as monkey [19], rat [20], and horse [21].…”