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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.

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Mesenchymal stem cell cultures display a typical MSC phenotype and can be induced to differentiate towards osteogenic, adipogenic and chondrogenic lineages. (A) Flow cytometric analysis of CD29, CD105, CD166, CD44 and CD45 expression by mesenchymal stem cell (MSC) cultures at third passage (n = 4). Images depicting MSC cultures, at third passage, differentiated down osteogenic, adipogenic and chondrogenic lineages. Osteogenic differentiation was visualized by the presence of high levels of alkaline phosphatase (B) and using immunofluorescent detection of Alkaline phosphatase (red)/nuclei (blue) (C). Chondrogenic differentiation was characterized by Alcian blue staining (D) and the immunofluorescent detection of aggrecan (red)/nuclei (blue) (E). Adipogenic differentiation was visualized by the accumulation of lipid-containing vacuoles which stain red with oil red O (F) and using immunofluorescent detection of lipoprotein lipase (red)/nuclei (blue) (G).
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fig01: Mesenchymal stem cell cultures display a typical MSC phenotype and can be induced to differentiate towards osteogenic, adipogenic and chondrogenic lineages. (A) Flow cytometric analysis of CD29, CD105, CD166, CD44 and CD45 expression by mesenchymal stem cell (MSC) cultures at third passage (n = 4). Images depicting MSC cultures, at third passage, differentiated down osteogenic, adipogenic and chondrogenic lineages. Osteogenic differentiation was visualized by the presence of high levels of alkaline phosphatase (B) and using immunofluorescent detection of Alkaline phosphatase (red)/nuclei (blue) (C). Chondrogenic differentiation was characterized by Alcian blue staining (D) and the immunofluorescent detection of aggrecan (red)/nuclei (blue) (E). Adipogenic differentiation was visualized by the accumulation of lipid-containing vacuoles which stain red with oil red O (F) and using immunofluorescent detection of lipoprotein lipase (red)/nuclei (blue) (G).

Mentions: Cells harvested from femoral shaft marrows displayed typical characteristics of MSCs in culture. Mononuclear cell cultures derived from femoral shaft bone marrows produced confluent adherent layers of elongated fibroblast-like cells in mesenchymal culture conditions. Mesenchymal stem cells were characterized by their immunophenotype and differentiation potential at third passage of culture. Using flow cytometric analyses, MSCs were found to be uniformly positive for the mesenchymal markers CD105, CD166, CD44, but negative for CD45 which is consistent with the known MSC phenotype and excludes contamination of cultures with haemopoietic cells [31]. In addition, MSCs were successfully induced to differentiate towards osteogenic, adipogenic and chondrogenic lineages using the methods described (Figure 1).


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)

Mesenchymal stem cell cultures display a typical MSC phenotype and can be induced to differentiate towards osteogenic, adipogenic and chondrogenic lineages. (A) Flow cytometric analysis of CD29, CD105, CD166, CD44 and CD45 expression by mesenchymal stem cell (MSC) cultures at third passage (n = 4). Images depicting MSC cultures, at third passage, differentiated down osteogenic, adipogenic and chondrogenic lineages. Osteogenic differentiation was visualized by the presence of high levels of alkaline phosphatase (B) and using immunofluorescent detection of Alkaline phosphatase (red)/nuclei (blue) (C). Chondrogenic differentiation was characterized by Alcian blue staining (D) and the immunofluorescent detection of aggrecan (red)/nuclei (blue) (E). Adipogenic differentiation was visualized by the accumulation of lipid-containing vacuoles which stain red with oil red O (F) and using immunofluorescent detection of lipoprotein lipase (red)/nuclei (blue) (G).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig01: Mesenchymal stem cell cultures display a typical MSC phenotype and can be induced to differentiate towards osteogenic, adipogenic and chondrogenic lineages. (A) Flow cytometric analysis of CD29, CD105, CD166, CD44 and CD45 expression by mesenchymal stem cell (MSC) cultures at third passage (n = 4). Images depicting MSC cultures, at third passage, differentiated down osteogenic, adipogenic and chondrogenic lineages. Osteogenic differentiation was visualized by the presence of high levels of alkaline phosphatase (B) and using immunofluorescent detection of Alkaline phosphatase (red)/nuclei (blue) (C). Chondrogenic differentiation was characterized by Alcian blue staining (D) and the immunofluorescent detection of aggrecan (red)/nuclei (blue) (E). Adipogenic differentiation was visualized by the accumulation of lipid-containing vacuoles which stain red with oil red O (F) and using immunofluorescent detection of lipoprotein lipase (red)/nuclei (blue) (G).
Mentions: Cells harvested from femoral shaft marrows displayed typical characteristics of MSCs in culture. Mononuclear cell cultures derived from femoral shaft bone marrows produced confluent adherent layers of elongated fibroblast-like cells in mesenchymal culture conditions. Mesenchymal stem cells were characterized by their immunophenotype and differentiation potential at third passage of culture. Using flow cytometric analyses, MSCs were found to be uniformly positive for the mesenchymal markers CD105, CD166, CD44, but negative for CD45 which is consistent with the known MSC phenotype and excludes contamination of cultures with haemopoietic cells [31]. In addition, MSCs were successfully induced to differentiate towards osteogenic, adipogenic and chondrogenic lineages using the methods described (Figure 1).

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