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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

Identification of TrkC or NT-3 gene transfection and neural differentiation of co-cultured TrkC-MSCs and NT-3-MSCs in the 3D GS scaffold. a, b MSCs at 2 days after infected with Ad-TrkC and Ad-NT-3 in vitro, respectively. Note that TrkC-positive (a) and NT-3-positive (b) MSCs are evident. c, d Bar charts show the percentage of Tju-1-positive (c) and Map2-positive (d) cells in the M, MN, MT, and MN + MT groups. Asterisk indicates statistical significance compared with the M group (*P < 0.05), pound sign indicates significance compared with the MN group (#P < 0.05), and ampersand indicates significance compared with the MT group (&P < 0.05). The data were presented as mean ± standard deviation (n = 5), and one-way analysis of variance with least significant difference test statistics was performed. e Cells in the 3D GS scaffold were immunostained by Tju-1 and Map2 antibodies 14 days after culture. Tju-1-positive and Map2-positive cells were observed in all groups. Scale bars = 20 μm. 3D, Three-dimensional; GS, Gelatin sponge; M, MSCs; Map2, Microtubule-associated protein 2; MN, NT-3-MSCs; MSCs, Mesenchymal stem cells; MT, TrkC-MSCs; NT-3, Neurotrophin-3; Tju-1, β-tubulin III; TrkC, Tropomyosin receptor kinase C
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Fig1: Identification of TrkC or NT-3 gene transfection and neural differentiation of co-cultured TrkC-MSCs and NT-3-MSCs in the 3D GS scaffold. a, b MSCs at 2 days after infected with Ad-TrkC and Ad-NT-3 in vitro, respectively. Note that TrkC-positive (a) and NT-3-positive (b) MSCs are evident. c, d Bar charts show the percentage of Tju-1-positive (c) and Map2-positive (d) cells in the M, MN, MT, and MN + MT groups. Asterisk indicates statistical significance compared with the M group (*P < 0.05), pound sign indicates significance compared with the MN group (#P < 0.05), and ampersand indicates significance compared with the MT group (&P < 0.05). The data were presented as mean ± standard deviation (n = 5), and one-way analysis of variance with least significant difference test statistics was performed. e Cells in the 3D GS scaffold were immunostained by Tju-1 and Map2 antibodies 14 days after culture. Tju-1-positive and Map2-positive cells were observed in all groups. Scale bars = 20 μm. 3D, Three-dimensional; GS, Gelatin sponge; M, MSCs; Map2, Microtubule-associated protein 2; MN, NT-3-MSCs; MSCs, Mesenchymal stem cells; MT, TrkC-MSCs; NT-3, Neurotrophin-3; Tju-1, β-tubulin III; TrkC, Tropomyosin receptor kinase C

Mentions: To examine the protein expression of TrkC and NT-3 in genetically modified MSCs before seeding into GS scaffolds, IFS was used. About 75 % of TrkC-MSCs had TrkC-positive staining (Fig. 1a), and about 70 % of NT-3-MSCs had NT-3-positive staining (Fig. 1b).Fig. 1


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)

Identification of TrkC or NT-3 gene transfection and neural differentiation of co-cultured TrkC-MSCs and NT-3-MSCs in the 3D GS scaffold. a, b MSCs at 2 days after infected with Ad-TrkC and Ad-NT-3 in vitro, respectively. Note that TrkC-positive (a) and NT-3-positive (b) MSCs are evident. c, d Bar charts show the percentage of Tju-1-positive (c) and Map2-positive (d) cells in the M, MN, MT, and MN + MT groups. Asterisk indicates statistical significance compared with the M group (*P < 0.05), pound sign indicates significance compared with the MN group (#P < 0.05), and ampersand indicates significance compared with the MT group (&P < 0.05). The data were presented as mean ± standard deviation (n = 5), and one-way analysis of variance with least significant difference test statistics was performed. e Cells in the 3D GS scaffold were immunostained by Tju-1 and Map2 antibodies 14 days after culture. Tju-1-positive and Map2-positive cells were observed in all groups. Scale bars = 20 μm. 3D, Three-dimensional; GS, Gelatin sponge; M, MSCs; Map2, Microtubule-associated protein 2; MN, NT-3-MSCs; MSCs, Mesenchymal stem cells; MT, TrkC-MSCs; NT-3, Neurotrophin-3; Tju-1, β-tubulin III; TrkC, Tropomyosin receptor kinase C
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4482203&req=5

Fig1: Identification of TrkC or NT-3 gene transfection and neural differentiation of co-cultured TrkC-MSCs and NT-3-MSCs in the 3D GS scaffold. a, b MSCs at 2 days after infected with Ad-TrkC and Ad-NT-3 in vitro, respectively. Note that TrkC-positive (a) and NT-3-positive (b) MSCs are evident. c, d Bar charts show the percentage of Tju-1-positive (c) and Map2-positive (d) cells in the M, MN, MT, and MN + MT groups. Asterisk indicates statistical significance compared with the M group (*P < 0.05), pound sign indicates significance compared with the MN group (#P < 0.05), and ampersand indicates significance compared with the MT group (&P < 0.05). The data were presented as mean ± standard deviation (n = 5), and one-way analysis of variance with least significant difference test statistics was performed. e Cells in the 3D GS scaffold were immunostained by Tju-1 and Map2 antibodies 14 days after culture. Tju-1-positive and Map2-positive cells were observed in all groups. Scale bars = 20 μm. 3D, Three-dimensional; GS, Gelatin sponge; M, MSCs; Map2, Microtubule-associated protein 2; MN, NT-3-MSCs; MSCs, Mesenchymal stem cells; MT, TrkC-MSCs; NT-3, Neurotrophin-3; Tju-1, β-tubulin III; TrkC, Tropomyosin receptor kinase C
Mentions: To examine the protein expression of TrkC and NT-3 in genetically modified MSCs before seeding into GS scaffolds, IFS was used. About 75 % of TrkC-MSCs had TrkC-positive staining (Fig. 1a), and about 70 % of NT-3-MSCs had NT-3-positive staining (Fig. 1b).Fig. 1

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