EGFP transgene: a useful tool to track transplanted bone marrow mononuclear cell contribution to peripheral remyelination

Abstract

Bone marrow mononuclear cells (BMMC) constitute a heterogeneous population with potential to promote tissue regeneration. For this reason, this cell fraction has recently become a therapeutic alternative to mesenchymal stem cells, as culture is not required and phenotypic transformations can be hence avoided. In this work, and in order to attain long-lasting cell labeling and study longer survival times, we used BMMC isolated from adult transgenic rats expressing GFP to reproduce our wild type model and evaluate their remyelination ability in a reversible model of Wallerian degeneration. RT-PCR and flow cytometry analysis confirmed that cells isolated from the transgenic strain exhibited similar expression levels of markers specific to multipotent progenitors (CD34, CD90 and CD105) and Schwann cells (MPZ, MBP, S100β and p75) compared to wild type BMMC. BMMC expressing GFP retained their migration capacity, arriving exclusively at the injured nerve. Most importantly, and as detected through long-lasting cell tracking, some of these BMMC settled in the demyelinated area, mingled with endogenous cells, underwent phenotypic changes and colocalized with Schwann cell markers MBP and S100β. Also worth highlighting, transgenic BMMC replicated wild type BMMC effects in terms of MBP organization and levels. On the basis of these findings, BMMC isolated from transgenic animals constitute a useful tool to evaluate their role in peripheral nervous system demyelination-remyelination and the underlying mechanisms.

Sciatic nerve regeneration after traumatic injury using magnetic targeted adipose-derived mesenchymal stem cells

“…Working with systemically transplanted bone marrow mononuclear cells (BMMC), a heterogeneous fraction including mesenchymal stem cells (MSC), our group demonstrated beneficial effects on sciatic nerve regeneration, together with prevention of hyperalgesia in a Wallerian degeneration (WD) model. [4,5] MSC are adult stem cells, originally described only in bone marrow, which possess self-renewal and multi-lineage differentiation potential. They were popularized in 1991 by Arnold Caplan, who reported their ability to give rise to bone and cartilage.…”

Potential Research Tool of Stem Cells from Human Exfoliated Deciduous Teeth: Lentiviral Bmi-1 Immortalization with EGFP Marker

“…The fluorescence can be seen and quantified by using noninvasive methods like a fluorescence microscope, confocal microscope, and flow cytometry. EGFP was generated by enhanced fluorescent signal, by labelling the target cells more explicitly and by adding to the range of applications [55]. In this study, we find some potential uses for SHED-Bmi1-EGFP, including the cytocompatibility assessment of biomaterial scaffolds as well as in vivo tracking (Figure 5).…”

Corneal Reconstruction with EGFP-Labelled Limbal Mesenchymal Stem Cells in a Rabbit Model of Limbal Stem Cell Deficiency

“…For reliable analysis of stem cell survival, growth, differentiation, and migration after transplantation, it is critical to track them in the host tissue [ 31 ]. Sánchez-Abarca et al demonstrated that after subconjunctival transplantation of green fluorescent protein-labeled human MSCs to mice, the labeled cells were localized in the stroma and the corneal epithelium [ 28 ].…”