Cell and Organ Transplantology. 2023; 11(2):122-129.
DOI: 10.22494/cot.v11i2.159

Multipotent mesenchymal stromal cells of various origin reduce reactive gliosis in the hippocampal CA1 area during acute ischemia-reperfusion of the rat brain

Konovalov S.¹, Moroz V.¹, Konovalova N.¹, Deryabina O.², Toporova O.^2,3, Tochylovsky A.⁴, Kordium V.^2,3

• ¹National Pirogov Memorial Medical University, Vinnytsya, Ukraine
  
• ²Institute of Genetic and Regenerative Medicine, M. D. Strazhesko National Scientific Center of Cardiology, Clinical and Regenerative Medicine, National Academy of Medical Sciences of Ukraine, Kyiv, Ukraine Kyiv, Ukraine
• ³Institute of Molecular Biology and Genetics, National Academy of Sciences of Ukraine, Kyiv, Ukraine
• ⁴BioTexCom LLC, Kyiv, Ukraine

Abstract

Neuroprotective therapy for acute cerebrovascular accident is directly aimed at preserving the neurons of the penumbra, but the regeneration of glia in the affected area is also important. Glial cells have fast reactivity and are very sensitive to ischemic cerebral injury. New experimental studies have demonstrated the successful use of multipotent mesenchymal stromal cells (MMSCs) in stroke to modulate microglial activation.
The purpose of the study was to investigate the effect of MMSCs of different origin, MMSC lysate and citicoline on glial components in the brain ischemia-reperfusion model in rats.
Methods. The experiments were conducted on 190 4-month-old male Wistar rats weighing 160-190 g. After modeling ischemia-reperfusion of the brain by bilateral 20-minute occlusion of the internal carotid arteries, the animals were intravenously injected with human umbilical cord Wharton’s jelly-derived MMSCs, human and rat adipose tissue-derived MMSCs at a dose of 1×10⁶ cells/animal. Other groups were injected with fetal rat fibroblasts 1×10⁶ cells/animal in 0.2 mL 0.9 % saline or lysate from umbilical cord Wharton’s jelly-derived MMSCs (0.2 mL/animal). Control animals were injected i.v. with 0.9 % saline. The last group of rats received a single dose of the reference drug citicoline at a dose of 250 mg/kg. On the 7^th and 14^th days, the area and fluorescence intensity of cells expressing markers of astrocytes (GFAP), microglia (Iba1), and oligodendrocytes (Rip) were quantitatively assessed on sections of the hippocampal CA1 area using immunohistochemical examination with confocal microscopy.
Results. On the 7^th and 14^th days after ischemia-reperfusion in rats, the fluorescence intensity of GFAP-positive astrocytes and Iba1-positive microglial cells increased, which indicated pronounced reactive astrogliosis and activation of microglia in the CA1 area of the hippocampus. At the same time, ischemia-reperfusion did not significantly affect the content of Rip-positive oligodendrocytes in brain slices. The use of all treatment options (transplantation of MMSCs of different origin, their lysate, or the reference drug citicoline) had a cytoprotective effect and reduced reactive astro- and microgliosis, both on the 7^th and 14^th days after the injury. The best result was demonstrated by the treatment with human umbilical cord Wharton’s jelly-derived MMSCs.
Conclusion. Brain ischemia-reperfusion induces reactive gliosis by activating GFAP-positive and Iba1-positive glial cells in all layers of the hippocampus. The use of human umbilical cord Wharton’s jelly-derived MMSCs and fetal rat fibroblasts significantly reduces its intensity both on the 7^th and on the 14^th days after modeling the ischemic brain injury.

Key words: ischemic brain injury; hippocampus; glia; neuroprotection; multipotent mesenchymal stromal cells; citicoline

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