Mesenchymal Stem Cell Conditioned Medium Increases Glial Reactivity and Decreases Neuronal Survival in Spinal Cord Slice Cultures

Overview

Journal Biochem Biophys Rep

Specialty Biochemistry

Date 2021 Mar 15

PMID 33718633

Citations 6

Authors

Chelsea R Wood

Esri H Juarez

Francesco Ferrini

Peter Myint

John Innes

Laura Lossi

Adalberto Merighi

William E B Johnson

Affiliations

Soon will be listed here.

Abstract

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 reduced inflammation and increased nerve regeneration but MSC survival was short-lived, highlighting that beneficial action may derive from the secretome. Previous 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 Ca transients per cell were significantly increased. These results contradict previous and 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.

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References

Menezes K, Nascimento M, Goncalves J, Cruz A, Lopes D, Curzio B . Human mesenchymal cells from adipose tissue deposit laminin and promote regeneration of injured spinal cord in rats. PLoS One. 2014; 9(5):e96020. PMC: 4022508. DOI: 10.1371/journal.pone.0096020. View

McMahill B, Borjesson D, Sieber-Blum M, Nolta J, Sturges B . Stem cells in canine spinal cord injury--promise for regenerative therapy in a large animal model of human disease. Stem Cell Rev Rep. 2014; 11(1):180-93. DOI: 10.1007/s12015-014-9553-9. View

Tan Y, Uchida K, Nakajima H, Guerrero A, Watanabe S, Hirai T . Blockade of interleukin 6 signaling improves the survival rate of transplanted bone marrow stromal cells and increases locomotor function in mice with spinal cord injury. J Neuropathol Exp Neurol. 2013; 72(10):980-93. DOI: 10.1097/NEN.0b013e3182a79de9. View

Patar A, Dockery P, Howard L, McMahon S . Cell viability in three ex vivo rat models of spinal cord injury. J Anat. 2018; 234(2):244-251. PMC: 6326831. DOI: 10.1111/joa.12909. View

Park H, Lim M, Jung H, Lee S, Paik K, Chang M . Human mesenchymal stem cell-derived Schwann cell-like cells exhibit neurotrophic effects, via distinct growth factor production, in a model of spinal cord injury. Glia. 2010; 58(9):1118-32. DOI: 10.1002/glia.20992. View

Pandamooz S, Salehi M, Zibaii M, Safari A, Nabiuni M, Ahmadiani A . Modeling traumatic injury in organotypic spinal cord slice culture obtained from adult rat. Tissue Cell. 2019; 56:90-97. DOI: 10.1016/j.tice.2019.01.002. View

Longair M, Baker D, Armstrong J . Simple Neurite Tracer: open source software for reconstruction, visualization and analysis of neuronal processes. Bioinformatics. 2011; 27(17):2453-4. DOI: 10.1093/bioinformatics/btr390. View

De Berdt P, Bottemanne P, Bianco J, Alhouayek M, Diogenes A, Lloyd A . Stem cells from human apical papilla decrease neuro-inflammation and stimulate oligodendrocyte progenitor differentiation via activin-A secretion. Cell Mol Life Sci. 2018; 75(15):2843-2856. PMC: 11105403. DOI: 10.1007/s00018-018-2764-5. View

Pandamooz S, Salehi M, Zibaii M, Ahmadiani A, Nabiuni M, Dargahi L . Epidermal neural crest stem cell-derived glia enhance neurotrophic elements in an ex vivo model of spinal cord injury. J Cell Biochem. 2017; 119(4):3486-3496. DOI: 10.1002/jcb.26520. View

10.

Park C, Kim K, Bae S, Son H, Myung P, Hong H . Cytokine secretion profiling of human mesenchymal stem cells by antibody array. Int J Stem Cells. 2014; 2(1):59-68. PMC: 4021795. DOI: 10.15283/ijsc.2009.2.1.59. View

11.

Hofstetter C, Schwarz E, Hess D, Widenfalk J, El Manira A, Prockop D . Marrow stromal cells form guiding strands in the injured spinal cord and promote recovery. Proc Natl Acad Sci U S A. 2002; 99(4):2199-204. PMC: 122342. DOI: 10.1073/pnas.042678299. View

12.

Cho J, Park H, Park S, Roh S, Kang S, Paik K . Transplantation of mesenchymal stem cells enhances axonal outgrowth and cell survival in an organotypic spinal cord slice culture. Neurosci Lett. 2009; 454(1):43-8. DOI: 10.1016/j.neulet.2009.02.024. View

13.

Jeffery N, Hamilton L, Granger N . Designing clinical trials in canine spinal cord injury as a model to translate successful laboratory interventions into clinical practice. Vet Rec. 2011; 168(4):102-7. DOI: 10.1136/vr.d475. View

14.

Tsai M, Liou D, Lin Y, Weng C, Huang M, Huang W . Attenuating Spinal Cord Injury by Conditioned Medium from Bone Marrow Mesenchymal Stem Cells. J Clin Med. 2018; 8(1). PMC: 6352201. DOI: 10.3390/jcm8010023. View

15.

Dribin L, Barrett J . Two components of conditioned medium increase neuritic outgrowth from rat spinal cord explants. J Neurosci Res. 1982; 8(2-3):271-80. DOI: 10.1002/jnr.490080217. View

16.

Maxwell D, Fyffe R, Brown A . Fine structure of normal and degenerating primary afferent boutons associated with characterized spinocervical tract neurons in the cat. Neuroscience. 1984; 12(1):151-63. DOI: 10.1016/0306-4522(84)90144-1. View

17.

Gomez T, Spitzer N . In vivo regulation of axon extension and pathfinding by growth-cone calcium transients. Nature. 1999; 397(6717):350-5. DOI: 10.1038/16927. View

18.

Lossi L, Merighi A . The Use of Rodent Platforms in Neuroscience Translational Research With Attention to the 3Rs Philosophy. Front Vet Sci. 2018; 5:164. PMC: 6060265. DOI: 10.3389/fvets.2018.00164. View

19.

Massoto T, Santos A, Ramalho B, Almeida F, Blanco Martinez A, Marques S . Mesenchymal stem cells and treadmill training enhance function and promote tissue preservation after spinal cord injury. Brain Res. 2019; 1726:146494. DOI: 10.1016/j.brainres.2019.146494. View

20.

Ravikumar M, Jain S, Miller R, Capadona J, Selkirk S . An organotypic spinal cord slice culture model to quantify neurodegeneration. J Neurosci Methods. 2012; 211(2):280-8. DOI: 10.1016/j.jneumeth.2012.09.004. View