Therapeutic Effects of hiPSC-Derived Glial and Neuronal Progenitor Cells-Conditioned Medium in Experimental Ischemic Stroke in Rats.

Diana Salikhova,Vladimir Chekhonin,Konstantin Midiber,Maria Nikitina,Georgy Leonov,Victoria Mokrousova,Elvira Cherkashova,Konstantin Yarygin,Andrey Elchaninov,Liudmila Mikhaleva,Daria Namestnikova,Dmitry Goldshtein,Natalia Bulatenco,Gevorg Akopyan,Ilya Gubskiy,Timur Fatkhudinov,Tatiana Bukharova,Andrey Makarov

doi:10.3390/ijms22094694

Abstract

Transplantation of various types of stem cells as a possible therapy for stroke has been tested for years, and the results are promising. Recent investigations have shown that the administration of the conditioned media obtained after stem cell cultivation can also be effective in the therapy of the central nervous system pathology (hypothesis of their paracrine action). The aim of this study was to evaluate the therapeutic effects of the conditioned medium of hiPSC-derived glial and neuronal progenitor cells in the rat middle cerebral artery occlusion model of the ischemic stroke. Secretory activity of the cultured neuronal and glial progenitor cells was evaluated by proteomic and immunosorbent-based approaches. Therapeutic effects were assessed by overall survival, neurologic deficit and infarct volume dynamics, as well as by the end-point values of the apoptosis- and inflammation-related gene expression levels, the extent of microglia/macrophage infiltration and the numbers of formed blood vessels in the affected area of the brain. As a result, 31% of the protein species discovered in glial progenitor cells-conditioned medium and 45% in neuronal progenitor cells-conditioned medium were cell type specific. The glial progenitor cell-conditioned media showed a higher content of neurotrophins (BDNF, GDNF, CNTF and NGF). We showed that intra-arterial administration of glial progenitor cells-conditioned medium promoted a faster decrease in neurological deficit compared to the control group, reduced microglia/macrophage infiltration, reduced expression of pro-apoptotic gene Bax and pro-inflammatory cytokine gene Tnf, increased expression of anti-inflammatory cytokine genes (Il4, Il10, Il13) and promoted the formation of blood vessels within the damaged area. None of these effects were exerted by the neuronal progenitor cell-conditioned media. The results indicate pronounced cytoprotective, anti-inflammatory and angiogenic properties of soluble factors secreted by glial progenitor cells.

Highlights

IntroductionThe acute ischemic damage to brain tissue causes focal and generalized neuronal death with diverse neurological sequelae [1]
Deterioration of brain function in ischemic stroke is overwhelming
The obtained human induced pluripotent stem cells (hiPSCs) were morphologically similar to human embryonic stem cells; displayed a high nuclear-cytoplasmic ratio; expressed pluripotency marker genes OCT4, NANOG and SOX2; and were immunopositive for OCT4 and NANOG transcription factors and SSEA4 and TRA-1-81 proteoglycans (Figure 1a)

Summary

Introduction

The acute ischemic damage to brain tissue causes focal and generalized neuronal death with diverse neurological sequelae [1]. The efficacy of treatment and rehabilitation of the patients after acute cerebrovascular episodes remains extremely low. In the absence of etiological treatment in the subacute and chronic phases of the disease, it is important to boost the rehabilitation of patients by stimulating nervous tissue repair [2]. New efficient maintenance therapies for cerebrovascular diseases are in great demand. Neurotrophins, cytokines and other regulatory molecules accumulated in the cell culture media can exert complex positive effects in various pathological conditions. The great potentiality of stem cell-conditioned media in the treatment of neurodegenerative and vascular diseases has been demonstrated in a number of studies [7,8,9]

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