Mesenchymal stem cell conditioned medium increases glial reactivity and decreases neuronal survival in spinal cord slice cultures
Ex vivo spinal cord slice cultures (SCSC) allow study of spinal cord circuitry, maintaining stimuli responses comparable to live animals. Previously, we have shown that mesenchymal stem/stromal cell (MSC) transplantation in vivo reduced inflammation and increased nerve regeneration but MSC survival...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2021-07-01
|
| Series: | Biochemistry and Biophysics Reports |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405580821000704 |
| _version_ | 1819069982793793536 |
|---|---|
| author | Chelsea R. Wood Esri H. Juárez Francesco Ferrini Peter Myint John Innes Laura Lossi Adalberto Merighi William E.B. Johnson |
| author_facet | Chelsea R. Wood Esri H. Juárez Francesco Ferrini Peter Myint John Innes Laura Lossi Adalberto Merighi William E.B. Johnson |
| author_sort | Chelsea R. Wood |
| collection | DOAJ |
| description | Ex vivo spinal cord slice cultures (SCSC) allow study of spinal cord circuitry, maintaining stimuli responses comparable to live animals. Previously, we have shown that mesenchymal stem/stromal cell (MSC) transplantation in vivo reduced inflammation and increased nerve regeneration but MSC survival was short-lived, highlighting that beneficial action may derive from the secretome. Previous in vitro studies of MSC conditioned medium (CM) have also shown increased neuronal growth. In this study, murine SCSC were cultured in canine MSC CM (harvested from the adipose tissue of excised inguinal fat) and cell phenotypes analysed via immunohistochemistry and confocal microscopy. SCSC in MSC CM displayed enhanced viability after propidium iodide staining. GFAP immunoreactivity was significantly increased in SCSC in MSC CM compared to controls, but with no change in proteoglycan (NG2) immunoreactivity. In contrast, culture in MSC CM significantly decreased the prevalence of βIII-tubulin immunoreactive neurites, whilst Ca2+ transients per cell were significantly increased. These ex vivo results contradict previous in vitro and in vivo reports of how MSC and their secretome may affect the microenvironment of the spinal cord after injury and highlight the importance of a careful comparison of the different experimental conditions used to assess the potential of cell therapies for the treatment of spinal cord injury. |
| first_indexed | 2024-12-21T16:58:42Z |
| format | Article |
| id | doaj.art-da6a749fa6c94e27bc9e67486e5e52b1 |
| institution | Directory Open Access Journal |
| issn | 2405-5808 |
| language | English |
| last_indexed | 2024-12-21T16:58:42Z |
| publishDate | 2021-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Biochemistry and Biophysics Reports |
| spelling | doaj.art-da6a749fa6c94e27bc9e67486e5e52b12022-12-21T18:56:41ZengElsevierBiochemistry and Biophysics Reports2405-58082021-07-0126100976Mesenchymal stem cell conditioned medium increases glial reactivity and decreases neuronal survival in spinal cord slice culturesChelsea R. Wood0Esri H. Juárez1Francesco Ferrini2Peter Myint3John Innes4Laura Lossi5Adalberto Merighi6William E.B. Johnson7Department of Biological Sciences, University of Chester, Parkgate Road, Chester, CH1 4BJ, UK; Corresponding author.Department of Veterinary Sciences, University of Turin, Largo Paolo Braccini 2, I-10095, Grugliasco, TO, ItalyDepartment of Veterinary Sciences, University of Turin, Largo Paolo Braccini 2, I-10095, Grugliasco, TO, Italy; Université Laval, Department of Psychiatry and Neuroscience, G1K 7P4, Québec, CanadaVeterinary Tissue Bank Ltd., No.1 The Long Barn, Brynkinalt Business Centre, Chirk, Wrexham, LL14 5NS, UKVeterinary Tissue Bank Ltd., No.1 The Long Barn, Brynkinalt Business Centre, Chirk, Wrexham, LL14 5NS, UKDepartment of Veterinary Sciences, University of Turin, Largo Paolo Braccini 2, I-10095, Grugliasco, TO, ItalyDepartment of Veterinary Sciences, University of Turin, Largo Paolo Braccini 2, I-10095, Grugliasco, TO, ItalyDepartment of Biological Sciences, University of Chester, Parkgate Road, Chester, CH1 4BJ, UKEx vivo spinal cord slice cultures (SCSC) allow study of spinal cord circuitry, maintaining stimuli responses comparable to live animals. Previously, we have shown that mesenchymal stem/stromal cell (MSC) transplantation in vivo reduced inflammation and increased nerve regeneration but MSC survival was short-lived, highlighting that beneficial action may derive from the secretome. Previous in vitro studies of MSC conditioned medium (CM) have also shown increased neuronal growth. In this study, murine SCSC were cultured in canine MSC CM (harvested from the adipose tissue of excised inguinal fat) and cell phenotypes analysed via immunohistochemistry and confocal microscopy. SCSC in MSC CM displayed enhanced viability after propidium iodide staining. GFAP immunoreactivity was significantly increased in SCSC in MSC CM compared to controls, but with no change in proteoglycan (NG2) immunoreactivity. In contrast, culture in MSC CM significantly decreased the prevalence of βIII-tubulin immunoreactive neurites, whilst Ca2+ transients per cell were significantly increased. These ex vivo results contradict previous in vitro and in vivo reports of how MSC and their secretome may affect the microenvironment of the spinal cord after injury and highlight the importance of a careful comparison of the different experimental conditions used to assess the potential of cell therapies for the treatment of spinal cord injury.http://www.sciencedirect.com/science/article/pii/S2405580821000704Mesenchymal stem/stromal cellSpinal cord slice culturesAstrocyte hypertrophyNeuronal survivalCalcium imaging |
| spellingShingle | Chelsea R. Wood Esri H. Juárez Francesco Ferrini Peter Myint John Innes Laura Lossi Adalberto Merighi William E.B. Johnson Mesenchymal stem cell conditioned medium increases glial reactivity and decreases neuronal survival in spinal cord slice cultures Biochemistry and Biophysics Reports Mesenchymal stem/stromal cell Spinal cord slice cultures Astrocyte hypertrophy Neuronal survival Calcium imaging |
| title | Mesenchymal stem cell conditioned medium increases glial reactivity and decreases neuronal survival in spinal cord slice cultures |
| title_full | Mesenchymal stem cell conditioned medium increases glial reactivity and decreases neuronal survival in spinal cord slice cultures |
| title_fullStr | Mesenchymal stem cell conditioned medium increases glial reactivity and decreases neuronal survival in spinal cord slice cultures |
| title_full_unstemmed | Mesenchymal stem cell conditioned medium increases glial reactivity and decreases neuronal survival in spinal cord slice cultures |
| title_short | Mesenchymal stem cell conditioned medium increases glial reactivity and decreases neuronal survival in spinal cord slice cultures |
| title_sort | mesenchymal stem cell conditioned medium increases glial reactivity and decreases neuronal survival in spinal cord slice cultures |
| topic | Mesenchymal stem/stromal cell Spinal cord slice cultures Astrocyte hypertrophy Neuronal survival Calcium imaging |
| url | http://www.sciencedirect.com/science/article/pii/S2405580821000704 |
| work_keys_str_mv | AT chelsearwood mesenchymalstemcellconditionedmediumincreasesglialreactivityanddecreasesneuronalsurvivalinspinalcordslicecultures AT esrihjuarez mesenchymalstemcellconditionedmediumincreasesglialreactivityanddecreasesneuronalsurvivalinspinalcordslicecultures AT francescoferrini mesenchymalstemcellconditionedmediumincreasesglialreactivityanddecreasesneuronalsurvivalinspinalcordslicecultures AT petermyint mesenchymalstemcellconditionedmediumincreasesglialreactivityanddecreasesneuronalsurvivalinspinalcordslicecultures AT johninnes mesenchymalstemcellconditionedmediumincreasesglialreactivityanddecreasesneuronalsurvivalinspinalcordslicecultures AT lauralossi mesenchymalstemcellconditionedmediumincreasesglialreactivityanddecreasesneuronalsurvivalinspinalcordslicecultures AT adalbertomerighi mesenchymalstemcellconditionedmediumincreasesglialreactivityanddecreasesneuronalsurvivalinspinalcordslicecultures AT williamebjohnson mesenchymalstemcellconditionedmediumincreasesglialreactivityanddecreasesneuronalsurvivalinspinalcordslicecultures |