Int J Stem Cells.  2018 Nov;11(2):177-186. 10.15283/ijsc18071.

Olig2-expressing Mesenchymal Stem Cells Enhance Functional Recovery after Contusive Spinal Cord Injury

• Park HW
• Oh S
• Lee KH
• Lee BH
• Chang MS

• KMID: 2429550
• DOI: http://doi.org/10.15283/ijsc18071

Abstract

BACKGROUND AND OBJECTIVES
Glial scarring and inflammation after spinal cord injury (SCI) interfere with neural regeneration and functional recovery due to the inhibitory microenvironment of the injured spinal cord. Stem cell transplantation can improve functional recovery in experimental models of SCI, but many obstacles to clinical application remain due to concerns regarding the effectiveness and safety of stem cell transplantation for SCI patients. In this study, we investigated the effects of transplantation of human mesenchymal stem cells (hMSCs) that were genetically modified to express Olig2 in a rat model of SCI.
METHODS
Bone marrow-derived hMSCs were genetically modified to express Olig2 and transplanted one week after the induction of contusive SCI in a rat model. Spinal cords were harvested 7 weeks after transplantation.
RESULTS
Transplantation of Olig2-expressing hMSCs significantly improved functional recovery in a rat model of contusive SCI model compared to the control hMSC-transplanted group. Transplantation of Olig2-expressing hMSCs also attenuated glial scar formation in spinal cord lesions. Immunohistochemical analysis showed that transplanted Olig2-expressing hMSCs were partially differentiated into Olig1-positive oligodendrocyte-like cells in spinal cords. Furthermore, NF-M-positive axons were more abundant in the Olig2-expressing hMSC-transplanted group than in the control hMSC-transplanted group.
CONCLUSIONS
We suggest that Olig2-expressing hMSCs are a safe and optimal cell source for treating SCI.

Keyword

Olig2; Mesenchymal stem cells; Spinal cord injury; Transplantation

MeSH Terms

Axons
Cicatrix
Humans
Inflammation
Mesenchymal Stromal Cells*
Models, Animal
Models, Theoretical
Regeneration
Spinal Cord Injuries*
Spinal Cord*
Stem Cell Transplantation
Transplantation

Figure

• 

  Fig. 1 Survival and distribution of Olig2-GFP-hMSCs in spinal cord lesion after contusive SCI. (A) Green fluorescent protein (GFP) and Olig2 expression in genetically modified hMSCs. The GFP-infected (GFP-hMSCs) and Olig2-GFP-infected (Olig2-GFP-hMSCs) hMSCs were immunostained with an anti-Olig2 (red) antibody. Colocalization of Oilg2 and GFP in the nucleus of Olig2-GFP-hMSCs is shown in the merged image (yellow). Scale bars: 100 μm. (B) Schematic diagram of the experimental design. (C) Immunohistochemistry of transplanted hMSCs in spinal cord lesion at 7 weeks after transplantation. Longitudinal spinal cord sections were immunostained for GFP at 7 weeks after transplantation of GFP-hMSCs and Olig2-GFP-hMSCs. Scale bars: 500 μm. The left side is rostral and the superior side is dorsal.

• 

  Fig. 2 Behavioral analysis of locomotor function. Effect of transplantation of Olig2-GFP-hMSCs on functional recovery after SCI. BBB locomotor scores of rats with SCI before and after transplantation of vehicle (n=5), GFP-hMSCs (n=5) and Olig2-GFP-hMSCs (n=6) were analyzed. Arrow indicates the time point when transplantation was performed. Data are shown as mean±SEM. *p<0.05 vs. vehicle transplant group, †p<0.05 vs. GFP-hMSC transplant group.

• 

  Fig. 3 Morphometry of spinal cord lesion after contusion SCI. Longitudinal spinal cord sections from the vehicle transplant group (A), GFP-hMSC transplant group (B), and Olig2-GFP-hMSC transplant group (C) were stained with hematoxylin and eosin (H&E) at 8 weeks after SCI. Histological analyses were conducted of longitudinal sections encompassing the lesion area. Dotted line outlines the lesion boundary. (D) The histograms represent measurements of the lesion area of vehicle transplant group, GFP-hMSC transplant group and Olig2-GFP-hMSC transplant group. (E) Correlations between BBB score 7 weeks post-injury and lesion size in the spinal cord tissue. Data are shown as mean±SEM. The results are representative of at least three rats in each group. *p<0.05 vs. vehicle transplant group, †p<0.05 vs. GFP-hMSC transplant group.

• 

  Fig. 4 Olig1 immunohistochemistry of the spinal cord lesion at 7 weeks after transplantation. Longitudinal spinal cord sections were immunostained for Olig1 and GFP 7 weeks after transplantation of vehicle (A), GFP-hMSCs (B) and Olig2-GFP-hMSCs (C). The right panel is magnified images of the boxed area. White arrows show colocalization of Olig1 to GFP-positive cells. Scale bars: 500 μm. In all images, the left side is rostral and the superior side is dorsal.

• 

  Fig. 5 NF-M immunohistochemistry of the spinal cord lesion at 7 weeks after transplantation. Longitudinal spinal cord sections were immunostained for NF-M and GFP 7 weeks after transplantation of vehicle (A), GFP-hMSCs (B), and Olig2-GFP-hMSCs (C). The right panel shows magnified images of the boxed area. White arrows show colocalization of NF-M to GFP-positive cells. Scale bars: 500 μm. In all images, the left side is rostral and the superior side is dorsal.

• 

  Fig. 6 Iba1 immunohistochemistry of the spinal cord lesion 7 weeks after transplantation. Longitudinal spinal cord sections were immunostained for Iba1 and GFP 7 weeks after transplantation of vehicle (A), GFP-hMSCs (B), and Olig2-GFP-hMSCs (C). The right panel shows magnified images of the boxed area. Scale bars: 500 μm. In all images, the left side is rostral and the superior side is dorsal.

• 

  Fig. 7 GFAP immunohistochemistry of the spinal cord lesion at 7 weeks after transplantation. Longitudinal spinal cord sections were immunostained for GFAP and GFP 7 weeks after transplantation of vehicle (A), GFP-hMSCs (B), and Olig2-GFP-hMSCs (C). The right panel shows magnified images of the boxed area. White arrows show colocalization of GFAP to GFP-positive cells. Scale bars: 500 μm. In all images, the left side is rostral and the superior side is dorsal.

Reference

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