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The Inability of Ex Vivo Expanded Mesenchymal Stem/Stromal Cells to Survive in Newborn Mice and to Induce Transplantation Tolerance

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Abstract

An encounter of the developing immune system with an antigen results in the induction of immunological areactivity to this antigen. In the case of transplantation antigens, the application of allogeneic hematopoietic cells induces a state of neonatal transplantation tolerance. This tolerance depends on the establishment of cellular chimerism, when allogeneic cells survive in the neonatally treated recipient. Since mesenchymal stem/stromal cells (MSCs) have been shown to have low immunogenicity and often survive in allogeneic recipients, we attempted to use these cells for induction of transplantation tolerance. Newborn (less than 24 h old) C57BL/6 mice were injected intraperitoneally with 5 × 106 adipose tissue-derived MSCs isolated from allogeneic donors and the fate and survival of these cells were monitored. The impact of MSC application on the proportion of cell populations of the immune system and immunological reactivity was assessed. In addition, the survival of skin allografts in neonatally treated recipients was tested. We found that in vitro expanded MSCs did not survive in neonatal recipients, and the living MSCs were not detected few days after their application. Furthermore, there were no significant changes in the proportion of individual immune cell populations including CD4+ cell lineages, but we detected an apparent shift to the production of Th1 cytokines IL-2 and IFN-γ in neonatally treated mice. However, skin allografts in the MSC-treated recipients were promptly rejected. These results therefore show that in vitro expanded MSCs do not survive in neonatal recipients, but induce a cytokine imbalance without induction of transplantation tolerance.

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Acknowledgements

We thank to Dr. Jan Prochazka (supported by Czech Science Foundation project No. 19-05076 S) from the Czech Centre for Phenogenomics, Institute of Molecular Genetics of the Czech Academy of Sciences, Vestec, Czech Republic, for providing us with mT/mG mice.

Funding

This work was supported grant No. 19-02290 S from the Czech Science Foundatuion, and by the Charles University programs SVV 260435 and 20604315 PROGRES Q43.

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Authors and Affiliations

  1. Department of Nanotoxicology and Molecular Epidemiology, Institute of Experimental Medicine of the Czech Academy of Sciences, 14220, Prague 4, Czech Republic

    Vladimir Holan, Barbora Echalar, Katerina Palacka, Jan Kossl, Pavla Bohacova, Bianka Porubská, Magdalena Krulova, Eliska Javorkova & Alena Zajicova

  2. Department of Cell Biology, Faculty of Science, Charles University, 12843, Prague 2, Czech Republic

    Vladimir Holan, Barbora Echalar, Katerina Palacka, Jan Kossl, Pavla Bohacova, Bianka Porubská, Magdalena Krulova & Eliska Javorkova

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  1. Vladimir Holan
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Contributions

All authors contributed to this study. VH, MK and AZ designed the study, VH, BE, KP, JK, PB. BP, MK, EJ, and AZ performed the experiments, VH and MK wrote the paper, all authors read and approved the final manuscript.

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Correspondence to Vladimir Holan.

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This study did not involve human participants. The experiments with animals were approved by the local Animal Ethics Committee of the Institute of Experimental Medicine of the Czech Academy of Sciences, Prague, and by ethical Committee of the Faculty of Science, Prague.

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Holan, V., Echalar, B., Palacka, K. et al. The Inability of Ex Vivo Expanded Mesenchymal Stem/Stromal Cells to Survive in Newborn Mice and to Induce Transplantation Tolerance. Stem Cell Rev and Rep 18, 2365–2375 (2022). https://doi.org/10.1007/s12015-022-10363-7

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