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Donor mesenchymal stem cell-derived neural-like cells transdifferentiate into myelin-forming cells and promote axon regeneration in rat spinal cord transection.

Qiu XC, Jin H, Zhang RY, Ding Y, Zeng X, Lai BQ, Ling EA, Wu JL, Zeng YS - Stem Cell Res Ther (2015)

Bottom Line: In the latter, the MSC-derived myelin-forming cells established myelin sheaths associated with the host regenerating axons.In addition, the cortical motor evoked potential and hindlimb locomotion were significantly ameliorated in the rat spinal cord transected in the MN + MT group compared with the GS and MSC groups.Grafted MSC-derived neural-like cells in the GS scaffold can transdifferentiate into myelin-forming cells in the completely transected rat spinal cord.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory for Stem Cells and Tissue Engineering (Sun Yat-sen University), Ministry of Education, Guangzhou, 510080, China. qiuxuecheng1990@163.com.

ABSTRACT

Introduction: Severe spinal cord injury often causes temporary or permanent damages in strength, sensation, or autonomic functions below the site of the injury. So far, there is still no effective treatment for spinal cord injury. Mesenchymal stem cells (MSCs) have been used to repair injured spinal cord as an effective strategy. However, the low neural differentiation frequency of MSCs has limited its application. The present study attempted to explore whether the grafted MSC-derived neural-like cells in a gelatin sponge (GS) scaffold could maintain neural features or transdifferentiate into myelin-forming cells in the transected spinal cord.

Methods: We constructed an engineered tissue by co-seeding of MSCs with genetically enhanced expression of neurotrophin-3 (NT-3) and its high-affinity receptor tropomyosin receptor kinase C (TrkC) separately into a three-dimensional GS scaffold to promote the MSCs differentiating into neural-like cells and transplanted it into the gap of a completely transected rat spinal cord. The rats received extensive post-operation care, including cyclosporin A administrated once daily for 2 months.

Results: MSCs modified genetically could differentiate into neural-like cells in the MN + MT (NT-3-MSCs + TrKC-MSCs) group 14 days after culture in the GS scaffold. However, after the MSC-derived neural-like cells were transplanted into the injury site of spinal cord, some of them appeared to lose the neural phenotypes and instead transdifferentiated into myelin-forming cells at 8 weeks. In the latter, the MSC-derived myelin-forming cells established myelin sheaths associated with the host regenerating axons. And the injured host neurons were rescued, and axon regeneration was induced by grafted MSCs modified genetically. In addition, the cortical motor evoked potential and hindlimb locomotion were significantly ameliorated in the rat spinal cord transected in the MN + MT group compared with the GS and MSC groups.

Conclusion: Grafted MSC-derived neural-like cells in the GS scaffold can transdifferentiate into myelin-forming cells in the completely transected rat spinal cord.

No MeSH data available.


Related in: MedlinePlus

Assessment of axonal regeneration in the injury/graft site of spinal cord at 8 weeks after cell transplantation. a The expression of neurofilament 200 (NF200) and chondroitin sulfate proteoglycans clone CS-56 (CSPG) in the gelatin sponge (GS), mesenchymal stem cells (MSCs), and NT-3-MSCs (MN) + TrkC-MSCs (MT) groups was detected by Western blotting (WB). b, c Bar charts demonstrate semi-quantitative analysis of the level of NF200 and CSPG expressions. In the MN + MT group, the level of NF200 expression was higher than in the GS and MSCs groups (b) (*P < 0.05, #P < 0.05). The level of CSPG expression was lower in the MN + MT and MSCs groups compared with the GS group (c) (*P < 0.05). d Bar charts demonstrate the axon density in the rostral, central, and caudal areas of the injury/graft site. The axon density in the rostral and central areas of the injury/graft site was higher in the MN + MT and MSCs groups compared with the GS group (*P < 0.05). The axon density in the caudal area of the injury/graft site in the MN + MT group was higher than in the MSCs and GS groups (*P < 0.05, #P < 0.05). e–g NF200-positive nerve fiber regeneration in the GS, MSCs, and MN + MT groups. The enlarged images from the rostral, central, and caudal areas of the injury/graft site are shown in E1–E3, F1–F3, and G1–G3. One-way analysis of variance with least significant difference test statistics was performed to compare the axon density. Scale bars = 50 μm
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Fig5: Assessment of axonal regeneration in the injury/graft site of spinal cord at 8 weeks after cell transplantation. a The expression of neurofilament 200 (NF200) and chondroitin sulfate proteoglycans clone CS-56 (CSPG) in the gelatin sponge (GS), mesenchymal stem cells (MSCs), and NT-3-MSCs (MN) + TrkC-MSCs (MT) groups was detected by Western blotting (WB). b, c Bar charts demonstrate semi-quantitative analysis of the level of NF200 and CSPG expressions. In the MN + MT group, the level of NF200 expression was higher than in the GS and MSCs groups (b) (*P < 0.05, #P < 0.05). The level of CSPG expression was lower in the MN + MT and MSCs groups compared with the GS group (c) (*P < 0.05). d Bar charts demonstrate the axon density in the rostral, central, and caudal areas of the injury/graft site. The axon density in the rostral and central areas of the injury/graft site was higher in the MN + MT and MSCs groups compared with the GS group (*P < 0.05). The axon density in the caudal area of the injury/graft site in the MN + MT group was higher than in the MSCs and GS groups (*P < 0.05, #P < 0.05). e–g NF200-positive nerve fiber regeneration in the GS, MSCs, and MN + MT groups. The enlarged images from the rostral, central, and caudal areas of the injury/graft site are shown in E1–E3, F1–F3, and G1–G3. One-way analysis of variance with least significant difference test statistics was performed to compare the axon density. Scale bars = 50 μm

Mentions: At 8 weeks after surgery, 12 rats of the GS, MSC, and MN + MT groups (n = 4 for each group) were sacrificed for WB analysis to evaluate axonal regeneration and glial scar formation. The NF200 and CSPG expression level in the injury/graft site of spinal cord was detected in three groups (Fig. 5a). The NF200 expression level was higher, and CSPG expression was lower, in the MSC and MN + MT groups compared with the GS group (Fig. 5b, c; P < 0.05).Fig. 5


Donor mesenchymal stem cell-derived neural-like cells transdifferentiate into myelin-forming cells and promote axon regeneration in rat spinal cord transection.

Qiu XC, Jin H, Zhang RY, Ding Y, Zeng X, Lai BQ, Ling EA, Wu JL, Zeng YS - Stem Cell Res Ther (2015)

Assessment of axonal regeneration in the injury/graft site of spinal cord at 8 weeks after cell transplantation. a The expression of neurofilament 200 (NF200) and chondroitin sulfate proteoglycans clone CS-56 (CSPG) in the gelatin sponge (GS), mesenchymal stem cells (MSCs), and NT-3-MSCs (MN) + TrkC-MSCs (MT) groups was detected by Western blotting (WB). b, c Bar charts demonstrate semi-quantitative analysis of the level of NF200 and CSPG expressions. In the MN + MT group, the level of NF200 expression was higher than in the GS and MSCs groups (b) (*P < 0.05, #P < 0.05). The level of CSPG expression was lower in the MN + MT and MSCs groups compared with the GS group (c) (*P < 0.05). d Bar charts demonstrate the axon density in the rostral, central, and caudal areas of the injury/graft site. The axon density in the rostral and central areas of the injury/graft site was higher in the MN + MT and MSCs groups compared with the GS group (*P < 0.05). The axon density in the caudal area of the injury/graft site in the MN + MT group was higher than in the MSCs and GS groups (*P < 0.05, #P < 0.05). e–g NF200-positive nerve fiber regeneration in the GS, MSCs, and MN + MT groups. The enlarged images from the rostral, central, and caudal areas of the injury/graft site are shown in E1–E3, F1–F3, and G1–G3. One-way analysis of variance with least significant difference test statistics was performed to compare the axon density. Scale bars = 50 μm
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Fig5: Assessment of axonal regeneration in the injury/graft site of spinal cord at 8 weeks after cell transplantation. a The expression of neurofilament 200 (NF200) and chondroitin sulfate proteoglycans clone CS-56 (CSPG) in the gelatin sponge (GS), mesenchymal stem cells (MSCs), and NT-3-MSCs (MN) + TrkC-MSCs (MT) groups was detected by Western blotting (WB). b, c Bar charts demonstrate semi-quantitative analysis of the level of NF200 and CSPG expressions. In the MN + MT group, the level of NF200 expression was higher than in the GS and MSCs groups (b) (*P < 0.05, #P < 0.05). The level of CSPG expression was lower in the MN + MT and MSCs groups compared with the GS group (c) (*P < 0.05). d Bar charts demonstrate the axon density in the rostral, central, and caudal areas of the injury/graft site. The axon density in the rostral and central areas of the injury/graft site was higher in the MN + MT and MSCs groups compared with the GS group (*P < 0.05). The axon density in the caudal area of the injury/graft site in the MN + MT group was higher than in the MSCs and GS groups (*P < 0.05, #P < 0.05). e–g NF200-positive nerve fiber regeneration in the GS, MSCs, and MN + MT groups. The enlarged images from the rostral, central, and caudal areas of the injury/graft site are shown in E1–E3, F1–F3, and G1–G3. One-way analysis of variance with least significant difference test statistics was performed to compare the axon density. Scale bars = 50 μm
Mentions: At 8 weeks after surgery, 12 rats of the GS, MSC, and MN + MT groups (n = 4 for each group) were sacrificed for WB analysis to evaluate axonal regeneration and glial scar formation. The NF200 and CSPG expression level in the injury/graft site of spinal cord was detected in three groups (Fig. 5a). The NF200 expression level was higher, and CSPG expression was lower, in the MSC and MN + MT groups compared with the GS group (Fig. 5b, c; P < 0.05).Fig. 5

Bottom Line: In the latter, the MSC-derived myelin-forming cells established myelin sheaths associated with the host regenerating axons.In addition, the cortical motor evoked potential and hindlimb locomotion were significantly ameliorated in the rat spinal cord transected in the MN + MT group compared with the GS and MSC groups.Grafted MSC-derived neural-like cells in the GS scaffold can transdifferentiate into myelin-forming cells in the completely transected rat spinal cord.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory for Stem Cells and Tissue Engineering (Sun Yat-sen University), Ministry of Education, Guangzhou, 510080, China. qiuxuecheng1990@163.com.

ABSTRACT

Introduction: Severe spinal cord injury often causes temporary or permanent damages in strength, sensation, or autonomic functions below the site of the injury. So far, there is still no effective treatment for spinal cord injury. Mesenchymal stem cells (MSCs) have been used to repair injured spinal cord as an effective strategy. However, the low neural differentiation frequency of MSCs has limited its application. The present study attempted to explore whether the grafted MSC-derived neural-like cells in a gelatin sponge (GS) scaffold could maintain neural features or transdifferentiate into myelin-forming cells in the transected spinal cord.

Methods: We constructed an engineered tissue by co-seeding of MSCs with genetically enhanced expression of neurotrophin-3 (NT-3) and its high-affinity receptor tropomyosin receptor kinase C (TrkC) separately into a three-dimensional GS scaffold to promote the MSCs differentiating into neural-like cells and transplanted it into the gap of a completely transected rat spinal cord. The rats received extensive post-operation care, including cyclosporin A administrated once daily for 2 months.

Results: MSCs modified genetically could differentiate into neural-like cells in the MN + MT (NT-3-MSCs + TrKC-MSCs) group 14 days after culture in the GS scaffold. However, after the MSC-derived neural-like cells were transplanted into the injury site of spinal cord, some of them appeared to lose the neural phenotypes and instead transdifferentiated into myelin-forming cells at 8 weeks. In the latter, the MSC-derived myelin-forming cells established myelin sheaths associated with the host regenerating axons. And the injured host neurons were rescued, and axon regeneration was induced by grafted MSCs modified genetically. In addition, the cortical motor evoked potential and hindlimb locomotion were significantly ameliorated in the rat spinal cord transected in the MN + MT group compared with the GS and MSC groups.

Conclusion: Grafted MSC-derived neural-like cells in the GS scaffold can transdifferentiate into myelin-forming cells in the completely transected rat spinal cord.

No MeSH data available.


Related in: MedlinePlus