ST2-Conditioned Medium Fosters Dorsal Horn Cell Excitability and Synaptic Transmission in Cultured Mouse Spinal Cord

Esri H Juárez, Chelsea Wood, Rebecca Davies, Oksana Kehoe, William E. B. Johnson, Adalberto Merighi, Francesco Ferrini

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Conditioned medium obtained from bone marrow-derived stem cells has been proposed as a novel cell-free therapy in spinal cord injury and neuropathic pain, yet the direct effect on spinal neuron function has never been investigated. Here, we adopted spinal cord organotypic cultures (SCOCs) as an experimental model to probe the effect of ST2 murine mesenchymal stem cells-conditioned medium (ST2-CM) on dorsal horn (DH) neuron functional properties. Three days of SCOC exposure to ST2-CM increased neuronal activity measured by Fos expression, as well as spontaneous or induced firing. We showed that the increase in neuronal excitability was associated with changes in both intrinsic membrane properties and an enhanced excitatory drive. The increased excitability at the single-cell level was substantiated at the network level by detecting synchronous bursts of calcium waves across DH neurons. Altogether, SCOCs represent a viable tool to probe mesenchymal cells' effect on intact neuronal networks. Our findings indicate that ST2-CM enhances neuronal activity and synaptic wiring in the spinal dorsal horn. Our data also support the trophic role of mesenchymal cells CM in maintaining network activity in spinal circuits.
Original languageEnglish
Pages (from-to)2918-2928
Number of pages11
JournalStem Cell Reviews and Reports
Issue number8
Early online date6 Sept 2023
Publication statusE-pub ahead of print - 6 Sept 2023

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This work was supported by the National Centre for the Replacement Refinement & Reduction of Animals in Research Crack It solution “Neuroinflammation and nociception in a dish” ( fund.


  • Mesenchymal stem/stromal cells
  • Dorsal horn neurons
  • Spinal cord organotypic cultures
  • Firing activity
  • Calcium imaging


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