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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

Detection of mesenchymal stem cell (MSC)-derived neuron-like cells in gelatin sponge (GS) scaffold 14 days after culture. a, b Scanning electron microscopy (SEM) shows differentiating MSCs bearing long and branched processes (a) (red arrows) on the surface of GS. One of the processes appears to make contact with another cell body (b) (red arrowheads). c Transmission electron microscopy shows a few synapse-like structures between two processes of differentiating MSCs (red arrowheads). d PSD95 expression was detected by Western blotting (1 = the M group, 2 = the MN group, 3 = the MT group, 4 = the MN + MT group). PSD95 level is highest in the MN + MT group compared with the M, MN, and MT groups (One-way analysis of variance with least significant difference test statistics was performed; *P < 0.05, #P < 0.05, &P < 0.05). e–h Expression of PSD95 and SYP was detected by immunofluorescence staining in the M, MN, MT, and MN + MT groups. All groups exhibited PSD95-positive cells, whereas SYP-positive cells were absent in the M and MN groups (white arrows). Scale bars = 20 μm in (e–h). M, MSCs; MN, NT-3-MSCs; MT, TrkC-MSCs
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Fig2: Detection of mesenchymal stem cell (MSC)-derived neuron-like cells in gelatin sponge (GS) scaffold 14 days after culture. a, b Scanning electron microscopy (SEM) shows differentiating MSCs bearing long and branched processes (a) (red arrows) on the surface of GS. One of the processes appears to make contact with another cell body (b) (red arrowheads). c Transmission electron microscopy shows a few synapse-like structures between two processes of differentiating MSCs (red arrowheads). d PSD95 expression was detected by Western blotting (1 = the M group, 2 = the MN group, 3 = the MT group, 4 = the MN + MT group). PSD95 level is highest in the MN + MT group compared with the M, MN, and MT groups (One-way analysis of variance with least significant difference test statistics was performed; *P < 0.05, #P < 0.05, &P < 0.05). e–h Expression of PSD95 and SYP was detected by immunofluorescence staining in the M, MN, MT, and MN + MT groups. All groups exhibited PSD95-positive cells, whereas SYP-positive cells were absent in the M and MN groups (white arrows). Scale bars = 20 μm in (e–h). M, MSCs; MN, NT-3-MSCs; MT, TrkC-MSCs

Mentions: At 14 days after culture, genetically modified MSCs in GS scaffold in the MN + MT group were observed under the SEM. The differentiating MSCs were spindle-shaped cells bearing long branched processes and showed a good adherence on the GS surface (Fig. 2a). Some synapse-like contacts were observed (Fig. 2b). By TEM, synapse-like structures between cell processes were observed. The contact sites exhibited presynaptic electron-dense, a synaptic cleft, and a distinct post-synaptic electron-dense in the MN + MT group (Fig. 2c). By WB analysis, PSD95 expression level was higher in the MN + MT group than in other groups (Fig. 2d; P < 0.05). Moreover, PSD95 and SYP co-expressing cells were found with IFS only in the MN + MT and MT groups (Fig. 2e-h). The results suggest that genetically modified MSCs exhibit a neuron-like cell phenotype, such as the occurrence of synapse-like structures and expression of synaptic protein in GS scaffold 14 days after culture.Fig. 2


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)

Detection of mesenchymal stem cell (MSC)-derived neuron-like cells in gelatin sponge (GS) scaffold 14 days after culture. a, b Scanning electron microscopy (SEM) shows differentiating MSCs bearing long and branched processes (a) (red arrows) on the surface of GS. One of the processes appears to make contact with another cell body (b) (red arrowheads). c Transmission electron microscopy shows a few synapse-like structures between two processes of differentiating MSCs (red arrowheads). d PSD95 expression was detected by Western blotting (1 = the M group, 2 = the MN group, 3 = the MT group, 4 = the MN + MT group). PSD95 level is highest in the MN + MT group compared with the M, MN, and MT groups (One-way analysis of variance with least significant difference test statistics was performed; *P < 0.05, #P < 0.05, &P < 0.05). e–h Expression of PSD95 and SYP was detected by immunofluorescence staining in the M, MN, MT, and MN + MT groups. All groups exhibited PSD95-positive cells, whereas SYP-positive cells were absent in the M and MN groups (white arrows). Scale bars = 20 μm in (e–h). M, MSCs; MN, NT-3-MSCs; MT, TrkC-MSCs
© Copyright Policy - open-access
Related In: Results  -  Collection

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Fig2: Detection of mesenchymal stem cell (MSC)-derived neuron-like cells in gelatin sponge (GS) scaffold 14 days after culture. a, b Scanning electron microscopy (SEM) shows differentiating MSCs bearing long and branched processes (a) (red arrows) on the surface of GS. One of the processes appears to make contact with another cell body (b) (red arrowheads). c Transmission electron microscopy shows a few synapse-like structures between two processes of differentiating MSCs (red arrowheads). d PSD95 expression was detected by Western blotting (1 = the M group, 2 = the MN group, 3 = the MT group, 4 = the MN + MT group). PSD95 level is highest in the MN + MT group compared with the M, MN, and MT groups (One-way analysis of variance with least significant difference test statistics was performed; *P < 0.05, #P < 0.05, &P < 0.05). e–h Expression of PSD95 and SYP was detected by immunofluorescence staining in the M, MN, MT, and MN + MT groups. All groups exhibited PSD95-positive cells, whereas SYP-positive cells were absent in the M and MN groups (white arrows). Scale bars = 20 μm in (e–h). M, MSCs; MN, NT-3-MSCs; MT, TrkC-MSCs
Mentions: At 14 days after culture, genetically modified MSCs in GS scaffold in the MN + MT group were observed under the SEM. The differentiating MSCs were spindle-shaped cells bearing long branched processes and showed a good adherence on the GS surface (Fig. 2a). Some synapse-like contacts were observed (Fig. 2b). By TEM, synapse-like structures between cell processes were observed. The contact sites exhibited presynaptic electron-dense, a synaptic cleft, and a distinct post-synaptic electron-dense in the MN + MT group (Fig. 2c). By WB analysis, PSD95 expression level was higher in the MN + MT group than in other groups (Fig. 2d; P < 0.05). Moreover, PSD95 and SYP co-expressing cells were found with IFS only in the MN + MT and MT groups (Fig. 2e-h). The results suggest that genetically modified MSCs exhibit a neuron-like cell phenotype, such as the occurrence of synapse-like structures and expression of synaptic protein in GS scaffold 14 days after culture.Fig. 2

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