Limits...
Fusion between human mesenchymal stem cells and rodent cerebellar Purkinje cells.

Kemp K, Gordon D, Wraith DC, Mallam E, Hartfield E, Uney J, Wilkins A, Scolding N - Neuropathol. Appl. Neurobiol. (2011)

Bottom Line: We found that fusion between MSCs and cerebellar neurons did occur in vitro and that the frequency of cellular fusion increased in the presence of TNF-alpha and/or IFN-gamma. we believe that this is the first paper to define fusion and heterokaryon formation between human MSCs and rodent cerebellar neurons in vivo.We have also demonstrated that fusion between these cell populations occurs in vitro.These findings indicate that MSCs may be potential therapeutic agents for cerebellar diseases, and other neuroinflammatory and neurodegenerative disorders.

View Article: PubMed Central - PubMed

Affiliation: Multiple Sclerosis and Stem Cell Group, Institute of Clinical Neurosciences, UK. kevin.kemp@bristol.ac.uk

ABSTRACT

Aims: we explored whether cellular fusion and heterokaryon formation between human and rodent cells in the cerebellum of mice occurs after intravenous injection of human bone marrow-derived mesenchymal stem cells (MSCs). The influence of central nervous system inflammation on this process was also assessed. In addition, we examined whether tumour necrosis factor (TNF)-alpha and interferon (IFN)-gamma, factors associated with inflammation, increase cellular fusion between human MSCs and rodent cerebellar neurons in vitro.

Methods and results: human MSCs were intravenously injected into mice with experimental autoimmune encephalomyelitis (EAE) and control mice. After 22 days, mouse Purkinje cells expressing human Golgi Zone were found within the Purkinje cell layer of the cerebellum, indicating that fusion and heterokaryon formation had occurred. The numbers of heterokaryons in the cerebellum were markedly increased in mice with EAE compared with control mice. Rodent cerebellar neuronal cells labelled with enhanced green fluorescent proteinin vitro were co-cultured with human bone marrow-derived MSCs in the presence of TNF-alpha and/or IFN-gamma to determine their influence on fusion events. We found that fusion between MSCs and cerebellar neurons did occur in vitro and that the frequency of cellular fusion increased in the presence of TNF-alpha and/or IFN-gamma.

Conclusions: we believe that this is the first paper to define fusion and heterokaryon formation between human MSCs and rodent cerebellar neurons in vivo. We have also demonstrated that fusion between these cell populations occurs in vitro. These findings indicate that MSCs may be potential therapeutic agents for cerebellar diseases, and other neuroinflammatory and neurodegenerative disorders.

Show MeSH

Related in: MedlinePlus

Human mesenchymal stem cells (MSCs) fuse with rat cerebellar enhanced green fluorescent protein (EGFP)-transduced neurons in vitro. Confocal images are of rat EGFP-transduced E18 cerebellar neuronal/human MSC co-cultures immunologically labelled with Hoescht nuclear stain (blue; a), EGFP (green; b), human Golgi Zone (red; c) and merged image (d). (A) Low power figure; arrows indicate dual stained EGFP/human Golgi Zone (red/green) cells; (B) high power figure; arrow head (human Golgi Zone positive/EGFP negative MSC), arrow head and star (human Golgi Zone negative/EGFP positive neuron), arrow (human Golgi Zone positive/EGFP positive fused MSC/neuronal cell). (C) An enlarged area of the dashed box in (B) showing a bi-nucleated human Golgi Zone positive/EGFP positive fused MSC/neuronal cell.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4150530&req=5

fig05: Human mesenchymal stem cells (MSCs) fuse with rat cerebellar enhanced green fluorescent protein (EGFP)-transduced neurons in vitro. Confocal images are of rat EGFP-transduced E18 cerebellar neuronal/human MSC co-cultures immunologically labelled with Hoescht nuclear stain (blue; a), EGFP (green; b), human Golgi Zone (red; c) and merged image (d). (A) Low power figure; arrows indicate dual stained EGFP/human Golgi Zone (red/green) cells; (B) high power figure; arrow head (human Golgi Zone positive/EGFP negative MSC), arrow head and star (human Golgi Zone negative/EGFP positive neuron), arrow (human Golgi Zone positive/EGFP positive fused MSC/neuronal cell). (C) An enlarged area of the dashed box in (B) showing a bi-nucleated human Golgi Zone positive/EGFP positive fused MSC/neuronal cell.

Mentions: After 15 days co-culture, fused cells were identified using immunofluorescence microscopy as EGFP+ve/human Golgi Zone+ve cells. Fused cells were found in all MSC/neuronal cell co-cultures (Figure 5). The majority of fused cells were shown to be bi-nucleate, although in a proportion of double positive cells only one nucleus was identifiable. No differences were seen in MSC numbers after the culture period between conditions (Figure 6B); however, there was a significant increase in neuronal survival evident when exposing neuronal/MSC cultures to both TNF-alpha and IFN-gamma when compared with base media alone (Figure 6A). The frequency of fusion events (demonstrated by double labelling with EGFP and human Golgi Zone) between the two cell populations was significantly increased by the addition of TNF-alpha and/or IFN-gamma to co-cultures when compared with the absence of cytokines (Figure 6C). Given the increase in neuronal numbers seen in co-cultures exposed to TNF-alpha and IFN-gamma, we calculated the percentage of EGFP positive cells expressing human Golgi Zone, and found that TNF-alpha and IFN-gamma increased the proportion of double labelled cells, indicating that this was not merely a function of increasing neuronal numbers (Figure 6D).


Fusion between human mesenchymal stem cells and rodent cerebellar Purkinje cells.

Kemp K, Gordon D, Wraith DC, Mallam E, Hartfield E, Uney J, Wilkins A, Scolding N - Neuropathol. Appl. Neurobiol. (2011)

Human mesenchymal stem cells (MSCs) fuse with rat cerebellar enhanced green fluorescent protein (EGFP)-transduced neurons in vitro. Confocal images are of rat EGFP-transduced E18 cerebellar neuronal/human MSC co-cultures immunologically labelled with Hoescht nuclear stain (blue; a), EGFP (green; b), human Golgi Zone (red; c) and merged image (d). (A) Low power figure; arrows indicate dual stained EGFP/human Golgi Zone (red/green) cells; (B) high power figure; arrow head (human Golgi Zone positive/EGFP negative MSC), arrow head and star (human Golgi Zone negative/EGFP positive neuron), arrow (human Golgi Zone positive/EGFP positive fused MSC/neuronal cell). (C) An enlarged area of the dashed box in (B) showing a bi-nucleated human Golgi Zone positive/EGFP positive fused MSC/neuronal cell.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4150530&req=5

fig05: Human mesenchymal stem cells (MSCs) fuse with rat cerebellar enhanced green fluorescent protein (EGFP)-transduced neurons in vitro. Confocal images are of rat EGFP-transduced E18 cerebellar neuronal/human MSC co-cultures immunologically labelled with Hoescht nuclear stain (blue; a), EGFP (green; b), human Golgi Zone (red; c) and merged image (d). (A) Low power figure; arrows indicate dual stained EGFP/human Golgi Zone (red/green) cells; (B) high power figure; arrow head (human Golgi Zone positive/EGFP negative MSC), arrow head and star (human Golgi Zone negative/EGFP positive neuron), arrow (human Golgi Zone positive/EGFP positive fused MSC/neuronal cell). (C) An enlarged area of the dashed box in (B) showing a bi-nucleated human Golgi Zone positive/EGFP positive fused MSC/neuronal cell.
Mentions: After 15 days co-culture, fused cells were identified using immunofluorescence microscopy as EGFP+ve/human Golgi Zone+ve cells. Fused cells were found in all MSC/neuronal cell co-cultures (Figure 5). The majority of fused cells were shown to be bi-nucleate, although in a proportion of double positive cells only one nucleus was identifiable. No differences were seen in MSC numbers after the culture period between conditions (Figure 6B); however, there was a significant increase in neuronal survival evident when exposing neuronal/MSC cultures to both TNF-alpha and IFN-gamma when compared with base media alone (Figure 6A). The frequency of fusion events (demonstrated by double labelling with EGFP and human Golgi Zone) between the two cell populations was significantly increased by the addition of TNF-alpha and/or IFN-gamma to co-cultures when compared with the absence of cytokines (Figure 6C). Given the increase in neuronal numbers seen in co-cultures exposed to TNF-alpha and IFN-gamma, we calculated the percentage of EGFP positive cells expressing human Golgi Zone, and found that TNF-alpha and IFN-gamma increased the proportion of double labelled cells, indicating that this was not merely a function of increasing neuronal numbers (Figure 6D).

Bottom Line: We found that fusion between MSCs and cerebellar neurons did occur in vitro and that the frequency of cellular fusion increased in the presence of TNF-alpha and/or IFN-gamma. we believe that this is the first paper to define fusion and heterokaryon formation between human MSCs and rodent cerebellar neurons in vivo.We have also demonstrated that fusion between these cell populations occurs in vitro.These findings indicate that MSCs may be potential therapeutic agents for cerebellar diseases, and other neuroinflammatory and neurodegenerative disorders.

View Article: PubMed Central - PubMed

Affiliation: Multiple Sclerosis and Stem Cell Group, Institute of Clinical Neurosciences, UK. kevin.kemp@bristol.ac.uk

ABSTRACT

Aims: we explored whether cellular fusion and heterokaryon formation between human and rodent cells in the cerebellum of mice occurs after intravenous injection of human bone marrow-derived mesenchymal stem cells (MSCs). The influence of central nervous system inflammation on this process was also assessed. In addition, we examined whether tumour necrosis factor (TNF)-alpha and interferon (IFN)-gamma, factors associated with inflammation, increase cellular fusion between human MSCs and rodent cerebellar neurons in vitro.

Methods and results: human MSCs were intravenously injected into mice with experimental autoimmune encephalomyelitis (EAE) and control mice. After 22 days, mouse Purkinje cells expressing human Golgi Zone were found within the Purkinje cell layer of the cerebellum, indicating that fusion and heterokaryon formation had occurred. The numbers of heterokaryons in the cerebellum were markedly increased in mice with EAE compared with control mice. Rodent cerebellar neuronal cells labelled with enhanced green fluorescent proteinin vitro were co-cultured with human bone marrow-derived MSCs in the presence of TNF-alpha and/or IFN-gamma to determine their influence on fusion events. We found that fusion between MSCs and cerebellar neurons did occur in vitro and that the frequency of cellular fusion increased in the presence of TNF-alpha and/or IFN-gamma.

Conclusions: we believe that this is the first paper to define fusion and heterokaryon formation between human MSCs and rodent cerebellar neurons in vivo. We have also demonstrated that fusion between these cell populations occurs in vitro. These findings indicate that MSCs may be potential therapeutic agents for cerebellar diseases, and other neuroinflammatory and neurodegenerative disorders.

Show MeSH
Related in: MedlinePlus