J Korean Med Sci.  2012 Jun;27(6):586-593. 10.3346/jkms.2012.27.6.586.

Fate of Transplanted Bone Marrow Derived Mesenchymal Stem Cells Following Spinal Cord Injury in Rats by Transplantation Routes

• Kang ES
• Ha KY
• Kim YH

• KMID: 1421609
• DOI: http://doi.org/10.3346/jkms.2012.27.6.586

Abstract

This research was performed to investigate the differences of the transplanted cells' survival and differentiation, and its efficacy according to the delivery routes following spinal cord injury. Allogenic mesenchymal stem cells (MSCs) were transplanted intravenously (IV group) or intralesionally (IL group) at post-injury 1 day in rats. Behavioral improvement, engraftment and differentiation of the transplanted cells and the expression of neurotrophic factors of the transplanted groups were analyzed and compared with those of the control group. At 6 weeks post-injury, the mean BBB motor scales in the control, IV and IL groups were 6.5 +/- 1.8, 11.1 +/- 2.1, and 8.5 +/- 2.8, respectively. Regardless of the delivery route, the MSCs transplantation following spinal cord injuries presented better behavioral improvement. The differentiations of the engrafted cells were different according to the delivery routes. The engrafted cells predominantly differentiated into astrocytes in the IV group and on the other hand, engrafted cells of the IL group demonstrated relatively even neural and glial differentiation. The expressions of neuronal growth factor were significantly higher in the IL group (mean relative optical density, 2.4 +/- 0.15) than those in the control (2.16 +/- 0.04) or IV group (1.7 +/- 0.23). Transplantation of MSCs in the early stage of spinal cord injury gives a significant clinical improvement. However, the fate of the transplanted MSCs and expression of neuronal growth factors are different along the transplantation route.

Keyword

Mesenchymal Stem Cells; Spinal Cord Injuries; Stem Cell Transplantation; Neuronal Differentiation; Neuroprotection

MeSH Terms

Animals
Behavior, Animal
Bone Marrow Cells/*cytology
Brain-Derived Neurotrophic Factor/metabolism
Cell Differentiation
Drug Administration Routes
Male
Mesenchymal Stem Cell Transplantation
Mesenchymal Stromal Cells/*cytology
Nerve Growth Factor/metabolism
Rats
Rats, Sprague-Dawley
Spinal Cord Injuries/*therapy
Transplantation, Homologous
Brain-Derived Neurotrophic Factor
Nerve Growth Factor

Figure

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  Fig. 1 Flow cytomertric analysis of cultured cells with CD 45, 73, and 29. The positive expression of CD 73 and CD 29, and negative expression of CD 45 indicate its mesenchymal stem cell lineage.

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  Fig. 2 Type II collagen expression of the transplanted cells were observed. In both transplanted groups, cells with a colocalization of PKH 26 and collagen II were not detected. (A) The IV transplanted MSCs do not express type II collagen. (B) Some type II collagen expression are noted in the IL group, however, no colocalization are found with PKH26 expression (collagen was tagged with green fluorescence, magnification × 400, scale bar 20 µm).

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  Fig. 3 Various expressions of neural and glial cell makers of engrafted MSCs. PKH26 positive cells were mainly found at the injured sites. (A) Neuronal differentiation of the transplanted MSCs (n = 4, two tissue samples and six fields in each sample). (B) Oligodendrocyte differentiation of the transplanted MSCs. (C) Astrocyte differentiation of the transplanted MSCs. Arrow indicates co-localization of PKH and GFAP expression. IV transplanted MSCs were mainly expressed the astrocyte differentiation. The proportion of neuronal and oligodendrocyte differentiation were lower than that of IL transplanted MSCs (magnification × 200, scale bar 50 µm).

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  Fig. 4 Expression of BDNF. Post-injury 1 week BDNF levels in the spinal cord tissues were measured. The relative optical densities in the control, IV and IL groups are 1.58 ± 0.22, 1.39 ± 0.35, 1.70 ± 0.2 respectively. The IL group shows slightly higher level compared to those in the control and IV groups. However, BDNF expression does not show any significant difference between the groups (n = 4, P > 0.05).

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  Fig. 5 Expression of NGF. Post-injury 1 week NGF levels in the spinal cord tissues were measured. The relative optical densities in the control, IV and IL groups are 2.16 ± 0.04, 1.70 ± 0.23, 2.41 ± 0.15 respectively. The IL group shows significantly higher level compared to those in the control and IV groups. And there is also significant difference between the groups (n = 4, P < 0.05).

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  Fig. 6 Open field locomotor assessment using Basso-Beattie-Bresnahan scale tested at every week after SCI. All subjects show a gradual improvement in hindlimb function during the 6 week observation period. Stastistical analysis indicated that BBB scales in the MSCs transplanted group were significantly higher than those in the control group (n = 8, P < 0.05). The rats in the IV group show the highest improvement at the last follow-up.

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