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Biocompatibility of intravitreal injection of human mesenchymal stem cells in immunocompetent rabbits

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Abstract

Purpose

To evaluate the feasibility, safety, and biocompatibility of intravitreal injection of human mesenchymal stem cells (MSCs) in immunocompetent pigmented rabbits.

Materials and methods

Thirty-two pigmented rabbits (24 females, 8 males; Chinchilla-New Zealand White) were divided into 8 groups of 4 animals. Commercially prepared human MSCs were injected (0.05 ml) into the post-lens vitreous of the right eyes. Groups 1 and 4 received isotonic medium (Ringer lactate-based), groups 2, 5, 7, and 8 received a low dose of 15 × 106 cells/ml. Groups 3 and 6 received a high dose of 30 × 106 cells/ml. Clinical signs were evaluated and scored before MSCs injection and weekly for 2 or 6 weeks. Animals were sacrificed at 2 or 6 weeks after injection. Eyes, liver, spleen, and gonads were assessed by histology and by fluorescent in situ hybridization to evaluate survival and extraocular migration of MSCs.

Results

There were no relevant clinical findings between control and MSC-injected rabbit eyes at any time point. There were also no relevant histological findings between control and MSC-injected rabbits related to ocular, liver, spleen, or gonad tissues modifications. MSCs survived intravitreally for at least 2 weeks after injection. Extraocular migration of MSCs was not detected.

Conclusions

MSCs are safe and well-tolerated when administered intravitreally at a dose of 15 × 106 cells/ml in pigmented rabbits. These findings enable future research to explore the intravitreal use of commercially prepared allogenic human MSCs in clinical trials of retinal diseases.

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Acknowledgements

This work was supported by grants from the Consejería de Educación de la Junta de Castilla y León (grant number VA118U14); and the Centro en Red de Medicina Regenerativa y Terapia Celular de la Junta de Castilla y León, Spain. The FISH technique was performed by the Cytogenetics Oncology Unit (University of Salamanca, Spain) following the protocol provided by the manufacturer. I Fernandez-Bueno, GK Srivastava and S Tabera-Bartolomé were supported by RETICS (RD12/0034/0001), Instituto de Salud Carlos III, Spain; Centro en Red de Medicina Regenerativa y Terapia Celular de la Junta de Castilla y Leon, Spain; and Red de Terapia Celular (RD12/0019/0036), Instituto de Salud Carlos III, Spain, respectively. Sonia Labrador Velandia and María L Alonso-Alonso were supported by the Consejería de Educación de la Junta de Castilla y León and the Fondo Social Europeo. This study was presented in part at The Association for Research in Vision and Ophthalmology (ARVO) Annual Meeting 2015, and at the European College of Veterinary Ophthalmologists (ECVO) Annual Scientific Meeting 2015.

Funding

This work was supported by grants from the Consejería de Educación de la Junta de Castilla y León (grant number VA118U14); and the Centro en Red de Medicina Regenerativa y Terapia Celular de la Junta de Castilla y León, Spain.

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Authors and Affiliations

  1. Instituto Universitario de Oftalmobiología Aplicada (IOBA), Retina Group, University of Valladolid, Campus Miguel Delibes, Paseo de Belén 17, 47011, Valladolid, Spain

    Sonia Labrador Velandia, Salvatore Di Lauro, Maria Luz Alonso-Alonso, Girish Kumar Srivastava, José Carlos Pastor & Ivan Fernandez-Bueno

  2. Department of Ophthalmology, Hospital Clínico Universitario de Valladolid, Valladolid, Spain

    Sonia Labrador Velandia, Salvatore Di Lauro & José Carlos Pastor

  3. UPC (Cell-production Unit), Instituto de Biología y Genética Molecular (IBGM), University of Valladolid and Centro Superior de Investigaciones Científicas (CSIC), Valladolid, Spain

    Soraya Tabera Bartolomé

  4. Centro en Red de Medicina Regenerativa y Terapia Celular de Castilla y León, Valladolid, Spain

    Girish Kumar Srivastava, José Carlos Pastor & Ivan Fernandez-Bueno

  5. Red Temática de Investigación Cooperativa en Salud (RETICS), Oftared, Instituto de Salud Carlos III, Valladolid, Spain

    Girish Kumar Srivastava, José Carlos Pastor & Ivan Fernandez-Bueno

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  1. Sonia Labrador Velandia
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  2. Salvatore Di Lauro
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  3. Maria Luz Alonso-Alonso
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  4. Soraya Tabera Bartolomé
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  5. Girish Kumar Srivastava
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  7. Ivan Fernandez-Bueno
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Correspondence to Ivan Fernandez-Bueno.

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All authors certify that they have no affiliations with or involvement in any organization or entity with any financial interest, or non-financial interest in the subject matter or materials discussed in this manuscript.

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All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted in agreement with European (Council Directive 2010/63/UE) and Spanish regulations (RD 53/2013).

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Labrador Velandia, S., Di Lauro, S., Alonso-Alonso, M.L. et al. Biocompatibility of intravitreal injection of human mesenchymal stem cells in immunocompetent rabbits. Graefes Arch Clin Exp Ophthalmol 256, 125–134 (2018). https://doi.org/10.1007/s00417-017-3842-3

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  • DOI: https://doi.org/10.1007/s00417-017-3842-3

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