Direct Conversion of Equine Adipose-Derived Stem Cells into Induced Neuronal Cells Is Enhanced in Three-Dimensional Culture

Abstract

The ability to culture neurons from horses may allow further investigation into equine neurological disorders. In this study, we demonstrate the generation of induced neuronal cells from equine adipose-derived stem cells (EADSCs) using a combination of lentiviral vector expression of the neuronal transcription factors Brn2, Ascl1, Myt1l (BAM) and NeuroD1 and a defined chemical induction medium, with βIII-tubulin-positive induced neuronal cells displaying a distinct neuronal morphology of rounded and compact cell bodies, extensive neurite outgrowth, and branching of processes. Furthermore, we investigated the effects of dimensionality on neuronal transdifferentiation, comparing conventional two-dimensional (2D) monolayer culture against three-dimensional (3D) culture on a porous polystyrene scaffold. Neuronal transdifferentiation was enhanced in 3D culture, with evenly distributed cells located on the surface and throughout the scaffold. Transdifferentiation efficiency was increased in 3D culture, with an increase in mean percent conversion of more than 100% compared to 2D culture. Additionally, induced neuronal cells were shown to transit through a Nestin-positive precursor state, with MAP2 and Synapsin 2 expression significantly increased in 3D culture. These findings will help to increase our understanding of equine neuropathogenesis, with prospective roles in disease modeling, drug screening, and cellular replacement for treatment of equine neurological disorders.

C1q/TNF-Related Protein 3 (CTRP-3) Deficiency of Adipocytes Affects White Adipose Tissue Mass but Not Systemic CTRP-3 Concentrations

“…The upregulation of Myt1l in mature adipocytes might be a compensatory mechanism during adipose tissue loss and in response to declining leptin levels. Since intra-abdominal adipose tissue represents a densely innervated tissue and since Myt1l activates the transdifferentiation of adipocyte precursor cells (APCs) into functional neurons [30,31], one could speculate that it is activated in order to compensate for adipose tissue loss. However, future mechanistic studies are needed to address this speculation properly.…”

The Proangiogenic Potential of Mesenchymal Stem Cells and Their Therapeutic Applications

“…Adult stem cells can be found in many tissues such as brain [27], liver [28], heart [29], lung [30], and adipose [31]. Adult stem cells are multipotent; they can self-renew and differentiate to all the cell types in their tissue environment and as well as other lineages such as cardiomyocytes [32], neurons [33], and endothelial cells [34].…”