Oligodendroglia Generated From Adult Rat Adipose Tissue by Direct Cell Conversion.

Lara Vellosillo,Jorge Pascual-Guerra,José Antonio Rodríguez-Navarro,Daniel González-Nieto,Maria Paz Muñoz,Luis Carlos Barrio,Maria Del Val Toledo Lobo,Carlos Luis Paíno

doi:10.3389/fcell.2022.741499

Abstract

Obtaining oligodendroglial cells from dispensable tissues would be of great interest for autologous or immunocompatible cell replacement therapy in demyelinating diseases, as well as for studying myelin-related pathologies or testing therapeutic approaches in culture. We evaluated the feasibility of generating oligodendrocyte precursor cells (OPCs) from adult rat adipose tissue by expressing genes encoding transcription factors involved in oligodendroglial development. Adipose-derived mesenchymal cells were lentivirally transduced with tetracycline-inducible Sox10, Olig2, Zfp536, and/or Nkx6.1 transgenes. Immunostaining with the OPC-specific O4 monoclonal antibody was used to mark oligodendroglial induction. O4- and myelin-associated glycoprotein (MAG)-positive cells emerged after 3 weeks when using the Sox10 + Olig2 + Zfp536 combination, followed in the ensuing weeks by GFAP-, O1 antigen-, p75NTR (low-affinity NGF receptor)-, and myelin proteins-positive cells. The O4+ cell population progressively expanded, eventually constituting more than 70% of cells in culture by 5 months. Sox10 transgene expression was essential for generating O4+ cells but was insufficient for inducing a full oligodendroglial phenotype. Converted cells required continuous transgene expression to maintain their glial phenotype. Some vestigial characteristics of mesenchymal cells were maintained after conversion. Growth factor withdrawal and triiodothyronine (T3) supplementation generated mature oligodendroglial phenotypes, while FBS supplementation produced GFAP+- and p75NTR+-rich cultures. Converted cells also showed functional characteristics of neural-derived OPCs, such as the expression of AMPA, NMDA, kainate, and dopaminergic receptors, as well as similar metabolic responses to differentiation-inducing drugs. When co-cultured with rat dorsal root ganglion neurons, the converted cells differentiated and ensheathed multiple axons. We propose that functional oligodendroglia can be efficiently generated from adult rat mesenchymal cells by direct phenotypic conversion.

Highlights

Oligodendrocyte transplantation has long been proposed to be a feasible strategy for repairing demyelinated lesions (Blakemore and Franklin, 1991; Archer et al, 1997; Wang et al, 2013), an approach that would be facilitated by the use of autologous or immunocompatible cells
Several procedures for obtaining expandable and myelinogenic oligodendrocytes from pluripotent stem cells (PSCs) in culture have been reported (Keirstead et al, 2005; Izrael et al, 2007; Hu et al, 2009; Najm et al, 2011; Douvaras et al, 2014; Douvaras and Fossati, 2015; Piao et al, 2015), including protocols that can promote the differentiation of oligodendrocytes from induced PSCs, themselves generated through the reprogramming of adult somatic cells (Wang et al, 2013; Douvaras et al, 2014; Douvaras and Fossati, 2015)
adipose-derived stromal cells (ADSCs) were transduced with tetracycline-inducible Sox10, Olig2, Zfp536, and/or Nkx6.1 (N) transgenes to test if the exogenous expression of the encoded transcription factors could induce the conversion of these adult rat cells into oligodendroglia

Summary

Introduction

Oligodendrocyte transplantation has long been proposed to be a feasible strategy for repairing demyelinated lesions (Blakemore and Franklin, 1991; Archer et al, 1997; Wang et al, 2013), an approach that would be facilitated by the use of autologous or immunocompatible cells. Some groups have proposed the rapid generation of oligodendroglia from human iPSCs via the exogenous expression of a combination of Sox, Olig, and Nkx6.2 (Ehrlich et al, 2017) or only Sox (Garcia-Leon et al, 2018). This process involves three steps, namely, the generation, selection, characterization, and the growing of iPSCs from cultures of adult somatic cells; deriving neural progenitor cells from the iPSCs; and using transduction to express genes encoding oligodendroglial transcription factors in these cells. In addition to the protracted time needed for cell reprogramming, the risk remains that, unless all cells are differentiated, residual PSCs might continue to proliferate and produce teratomas

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Conclusion