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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

Myelin sheath formation in the injury/graft site of spinal cord at 8 weeks after mesenchymal stem cell (MSC)-derived neural-like cell transplantation. a–d Co-localization of green fluorescent protein (GFP) and myelin basic protein (MBP) in an MSC-derived neural-like cell as observed under a confocal microscope. The ortho section showed that the grafted GFP-positive cell was immunostained by MBP antibody and encircled the host neurofilament 200 (NF200)-positive axon (white arrowheads). e, f A myelin sheath was formed by a GFP-positive cell under the GFP-immunoelectron microscope observation. The red box in (e) was magnified in (f). f Red arrows indicated GFP-positive electron-dense 3, 3′-diaminobenzidine (DAB) deposited on the myelin sheath, and red arrowheads indicated GFP-positive electron-dense DAB deposited on the surface of nucleus. Asterisk indicates the nucleus of MSC-derived myelin-forming cell (f). g A GFP-positive cell deposited with DAB and counterstained by toluidine blue formed myelin-like structure (red arrowheads) in semithin section. Asterisk indicates DAB-positive cell body (g). Scale bars = 20 μm (a–d)
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Fig4: Myelin sheath formation in the injury/graft site of spinal cord at 8 weeks after mesenchymal stem cell (MSC)-derived neural-like cell transplantation. a–d Co-localization of green fluorescent protein (GFP) and myelin basic protein (MBP) in an MSC-derived neural-like cell as observed under a confocal microscope. The ortho section showed that the grafted GFP-positive cell was immunostained by MBP antibody and encircled the host neurofilament 200 (NF200)-positive axon (white arrowheads). e, f A myelin sheath was formed by a GFP-positive cell under the GFP-immunoelectron microscope observation. The red box in (e) was magnified in (f). f Red arrows indicated GFP-positive electron-dense 3, 3′-diaminobenzidine (DAB) deposited on the myelin sheath, and red arrowheads indicated GFP-positive electron-dense DAB deposited on the surface of nucleus. Asterisk indicates the nucleus of MSC-derived myelin-forming cell (f). g A GFP-positive cell deposited with DAB and counterstained by toluidine blue formed myelin-like structure (red arrowheads) in semithin section. Asterisk indicates DAB-positive cell body (g). Scale bars = 20 μm (a–d)

Mentions: To investigate whether the APC and MBP expressing cells grafted could differentiate into mature oligodendrocytes, two techniques—namely, the triple IFS and GFP-immunoelectron microscope (GFP-IEM)—were used to detect myelination in the injury/graft site. Very strikingly, the tubular and ellipsoidal myelin-like structures, which were formed by the GFP-positive cells grafted, were observed by the triple IFS in the injury/graft site of spinal cord in all rats of the MN + MT group (Fig. 4a–d). Myelin-like structure showing coexistence appeared to encircle the host NF200-positive nerve fiber (Fig. 4a–d). To further verify that grafted MSCs could differentiate into myelinating cells, we scrutinized the GFP-positive cells by the GFP-IEM. The results showed that GFP-positive cells were identified in the injury/graft site and they appeared to enwrap a few host axons to form myelin structures. GFP reaction products of electron-dense DAB staining were obviously localized in the cytoplasm near or associated with the cell nucleus in a myelin-forming cell (Fig. 4e, f). Moreover, the myelin-like structures with DAB reaction products of GFP were found in the injury/graft site in the semi-thin section counterstained by toluidine blue (Fig. 4g). The results suggest that the differentiating MSCs grafted can transdifferentiate into myelin-forming cells in the injury/graft site of spinal cord transected completely at 8 weeks post-transplantation.Fig. 4


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)

Myelin sheath formation in the injury/graft site of spinal cord at 8 weeks after mesenchymal stem cell (MSC)-derived neural-like cell transplantation. a–d Co-localization of green fluorescent protein (GFP) and myelin basic protein (MBP) in an MSC-derived neural-like cell as observed under a confocal microscope. The ortho section showed that the grafted GFP-positive cell was immunostained by MBP antibody and encircled the host neurofilament 200 (NF200)-positive axon (white arrowheads). e, f A myelin sheath was formed by a GFP-positive cell under the GFP-immunoelectron microscope observation. The red box in (e) was magnified in (f). f Red arrows indicated GFP-positive electron-dense 3, 3′-diaminobenzidine (DAB) deposited on the myelin sheath, and red arrowheads indicated GFP-positive electron-dense DAB deposited on the surface of nucleus. Asterisk indicates the nucleus of MSC-derived myelin-forming cell (f). g A GFP-positive cell deposited with DAB and counterstained by toluidine blue formed myelin-like structure (red arrowheads) in semithin section. Asterisk indicates DAB-positive cell body (g). Scale bars = 20 μm (a–d)
© Copyright Policy - open-access
Related In: Results  -  Collection

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Fig4: Myelin sheath formation in the injury/graft site of spinal cord at 8 weeks after mesenchymal stem cell (MSC)-derived neural-like cell transplantation. a–d Co-localization of green fluorescent protein (GFP) and myelin basic protein (MBP) in an MSC-derived neural-like cell as observed under a confocal microscope. The ortho section showed that the grafted GFP-positive cell was immunostained by MBP antibody and encircled the host neurofilament 200 (NF200)-positive axon (white arrowheads). e, f A myelin sheath was formed by a GFP-positive cell under the GFP-immunoelectron microscope observation. The red box in (e) was magnified in (f). f Red arrows indicated GFP-positive electron-dense 3, 3′-diaminobenzidine (DAB) deposited on the myelin sheath, and red arrowheads indicated GFP-positive electron-dense DAB deposited on the surface of nucleus. Asterisk indicates the nucleus of MSC-derived myelin-forming cell (f). g A GFP-positive cell deposited with DAB and counterstained by toluidine blue formed myelin-like structure (red arrowheads) in semithin section. Asterisk indicates DAB-positive cell body (g). Scale bars = 20 μm (a–d)
Mentions: To investigate whether the APC and MBP expressing cells grafted could differentiate into mature oligodendrocytes, two techniques—namely, the triple IFS and GFP-immunoelectron microscope (GFP-IEM)—were used to detect myelination in the injury/graft site. Very strikingly, the tubular and ellipsoidal myelin-like structures, which were formed by the GFP-positive cells grafted, were observed by the triple IFS in the injury/graft site of spinal cord in all rats of the MN + MT group (Fig. 4a–d). Myelin-like structure showing coexistence appeared to encircle the host NF200-positive nerve fiber (Fig. 4a–d). To further verify that grafted MSCs could differentiate into myelinating cells, we scrutinized the GFP-positive cells by the GFP-IEM. The results showed that GFP-positive cells were identified in the injury/graft site and they appeared to enwrap a few host axons to form myelin structures. GFP reaction products of electron-dense DAB staining were obviously localized in the cytoplasm near or associated with the cell nucleus in a myelin-forming cell (Fig. 4e, f). Moreover, the myelin-like structures with DAB reaction products of GFP were found in the injury/graft site in the semi-thin section counterstained by toluidine blue (Fig. 4g). The results suggest that the differentiating MSCs grafted can transdifferentiate into myelin-forming cells in the injury/graft site of spinal cord transected completely at 8 weeks post-transplantation.Fig. 4

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