Transgenic Research

, Volume 27, Issue 2, pp 135–153 | Cite as

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

  • Gonzalo Piñero
  • Vanina Usach
  • Paula A. Soto
  • Paula V. Monje
  • Patricia Setton-Avruj
Original Paper


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 p75NTR) 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.


Bone marrow mononuclear cells Transplantation Sciatic nerve injury Remyelination 



Wild type Wistar rat


Hemizygous EGFP transgenic strain of the Wistar rat [Wistar-TgN(CAG-GFP)184ys]


Bone marrow mononuclear cells isolated from WtWistar rat


Bone marrow mononuclear cells isolated from EGFPWistar rat



Ms. María Marta Rancez for her assistance in the elaboration of this manuscript. Ms. Marianella Vence for her valuable help in animal breeding and care. Mr. Roberto Fernandez, BS. Confocal microscopy service, IFIBYNE-CONICET-UBA. Microscopy National System (SNM), Ministerio de Ciencia, Tecnología e Innovación Productiva de la República Argentina. Biochem Pharm Plácida Baz and Biochem Ariel Billordo. Flow cytometry service, Immunogenetics laboratory, INGEM-CONICET-UBA. Flow Cytometry National Service (SNCF), Ministerio de Ciencia, Tecnología e Innovación Productiva de la República Argentina. This work was supported by Universidad de Buenos Aires (UBACYT 20020100101017) and CONICET/Ministerio de Ciencia, Tecnología e Innovación Productiva de la República Argentina (PIP 830 and PIP 567). The authors declare that they have no conflicts of interest with the contents of this article.

Supplementary material

11248_2018_62_MOESM1_ESM.tif (2.7 mb)
Suppl. Figure 1 Flow cytometry analysis of freshly isolated bone marrow cells and BMMC from WtWistar and EGFPWistar (n = 4). Gating strategy used to exclude debris (FSC-A vs. SSC-A), doublets (FSC-A vs. FSC-H) and dead cells (viable dye Zombie yellow™), in (a) WtWistar and (b) EGFPWistar total bone marrow cells, and the BMMC fraction from (c) wild type and (d) transgenic strain (TIFF 2753 kb)
11248_2018_62_MOESM2_ESM.tif (464 kb)
Suppl. Figure 2 Migration of transplanted EGFPBMMC (n = 3). Representative confocal microscopy images (40x) of naïve and sham nerves at 7 and 14 dpi. Nuclei were stained with Hoechst (blue) (TIFF 464 kb)
11248_2018_62_MOESM3_ESM.tif (2.8 mb)
Suppl. Figure 3 Effect of transplanted EGFPBMMC on MBP organization in non injured nerves (n = 5). Representative epifluorescence microscopy images of MBP immunoreaction (red) in a naïve and contralateral nerve and the proximal stump of EGFPBMMC-transplanted animals at 7 and 14 dpi (20x). Nuclei were stained with Hoechst (blue). Bar graphs showing IOD proportions in non treated (white) and EGFPBMMC-transplanted animals (green) at 7 and 14 dpi (AU, arbitrary units: MBP IOD relative to naïve nerves. Values expressed as the mean ± SD). Statistical analysis done through two-way ANOVA followed by Bonferroni’s multiple comparison post test (ns = not significant; * = p < 0.05; ** p < 0.01; *** = p < 0.001) (TIFF 2897 kb)


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Copyright information

© Springer International Publishing AG, part of Springer Nature 2018

Authors and Affiliations

  1. 1.CONICET, Instituto de Química y Fisicoquímica Biológicas Prof. Alejandro C. Paladini (IQUIFIB)Universidad de Buenos AiresBuenos AiresArgentina
  2. 2.The Miami Project to Cure Paralysis and the Department of Neurological SurgeryUniversity of Miami Miller School of MedicineMiamiUSA

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