Safety and Biodistribution of Human Bone Marrow-Derived Mesenchymal Stromal Cells Injected Intrathecally in Non-Obese Diabetic Severe Combined Immunodeficiency Mice: Preclinical Study



Mesenchymal stromal cells (MSCs) have potent immunomodulatory and neuroprotective properties, and have been tested in neurodegenerative diseases resulting in meaningful clinical improvements. Regulatory guidelines specify the need to perform preclinical studies prior any clinical trial, including biodistribution assays and tumourigenesis exclusion. We conducted a preclinical study of human bone marrow MSCs (hBM-MSCs) injected by intrathecal route in Non-Obese Diabetic Severe Combined Immunodeficiency mice, to explore cellular biodistribution and toxicity as a privileged administration method for cell therapy in Friedreich’s Ataxia.


For this purpose, 3 × 105 cells were injected by intrathecal route in 12 animals (experimental group) and the same volume of culture media in 6 animals (control group). Blood samples were collected at 24 h (n = 9) or 4 months (n = 9) to assess toxicity, and nine organs were harvested for histology and safety studies. Genomic DNA was isolated from all tissues, and mouse GAPDH and human β2M and β-actin genes were amplified by qPCR to analyze hBM-MSCs biodistribution.


There were no deaths nor acute or chronic toxicity. Hematology, biochemistry and body weight were in the range of normal values in all groups. At 24 h hBM-MSCs were detected in 4/6 spinal cords and 1/6 hearts, and at 4 months in 3/6 hearts and 1/6 brains of transplanted mice. No tumours were found.


This study demonstrated that intrathecal injection of hBM-MSCs is safe, non toxic and do not produce tumors. These results provide further evidence that hBM-MSCs might be used in a clinical trial in patients with FRDA.

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The authors acknowledge Dra. Carmen Algueró for assistant with flow cytometry, Dr. Tornel-Osorio (Hospital Clínico Universitario Virgen de la Arrixaca) for assistant with the hematology and biochemistry analysis and Dra. Antón-García (Genomic Lab, IMIB-Arrixaca) for technical assistance. This work was supported by the Fundación Mutua Madrileña (AP162842016), Asociación Granadina de Ataxia de Friederich (ASOGAF), Instituto de Salud Carlos III (ISCIII) Spanish Net of Cell Therapy (TerCel), RETICS subprogram of the I + D + I 2013–2016 Spanish National Plan, Projects “RD12/0019/0001”, “RD12/0019/0023”, “RD16/0011/0001” and “RD16/0011/0010” funded by ISCIII and co-founded by European Regional Development Funds.

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Correspondence to Mari Paz Quesada.

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Ethical statement

The use of human bone marrow cells was in accordance with the guidelines and regulations of the Ethics Committee of the Hospital Clínico Universitario Virgen de la Arrixaca (Murcia, Spain). All of the donors provided written informed consent prior to participation in this study. The procedures performed in this work involving animals were approved by the Ethical Committee on Animal Experimentation at University of Murcia (220/2016). All the experimental procedures involving animals were conducted in accordance with the Institutional Animal care guidelines of Murcia University.

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Quesada, M.P., García-Bernal, D., Pastor, D. et al. Safety and Biodistribution of Human Bone Marrow-Derived Mesenchymal Stromal Cells Injected Intrathecally in Non-Obese Diabetic Severe Combined Immunodeficiency Mice: Preclinical Study. Tissue Eng Regen Med 16, 525–538 (2019).

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  • Intrathecal transplantation
  • Bone marrow-derived mesenchymal stromal cells
  • Friedreich’s Ataxia
  • Neuroprotection
  • Preclinical study