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A 3D construct based on mesenchymal stromal cells, collagen microspheres and plasma clot supports the survival, proliferation and differentiation of hematopoietic cells in vivo

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Abstract

The hematopoietic niche is a specialized microenvironment that supports the survival, proliferation and differentiation of hematopoietic stem progenitor cells (HSPCs). Three-dimensional (3D) models mimicking hematopoiesis might allow in vitro and in vivo studies of the hematopoietic (HP) process. Here, we investigate the capacity of a 3D construct based on non-adherent murine bone marrow mononuclear cells (NA-BMMNCs), mesenchymal stromal cells (MSCs) and collagen microspheres (CMs), all embedded into plasma clot (PC) to support in vitro and in vivo hematopoiesis. Confocal analysis of the 3D hematopoietic construct (3D-HPC), cultured for 24 h, showed MSC lining the CM and the NA-BMMNCs closely associated with MSC. In vivo hematopoiesis was examined in 3D-HPC subcutaneously implanted in mice and harvested at different intervals. Hematopoiesis in the 3D-HPC was evaluated by histology, cell morphology, flow cytometry, confocal microscopy and hematopoietic colony formation assay. 3D-HPC implants were integrated and vascularized in the host tissue, after 3 months of implantation. Histological studies showed the presence of hematopoietic tissue with the presence of mature blood cells. Cells from 3D-HPC showed viability greater than 90%, expressed HSPCs markers, and formed hematopoietic colonies, in vitro. Confocal microscopy studies showed that MSCs adhered to the CM and NA-BMMNCs were scattered across the 3D-HPC area and in close association with MSC. In conclusion, the 3D-HPC mimics a hematopoietic niche supporting the survival, proliferation and differentiation of HSPCs, in vivo. 3D-HPC may allow evaluation of regulatory mechanisms involved in hematopoiesis.

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Acknowledgments

The authors wish to thank Patricia Rodriguez and Blau Laboratory for technical assistance. The authors wish to thank the support of Esal and Tresta.

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Author notes

  1. Carlos Bello-Rodriguez and Olga Wittig contributed equally to this work.

Authors and Affiliations

  1. Unidad de Terapia Celular - Laboratorio de Patología Celular y Molecular, Centro de Medicina Experimental, Instituto Venezolano de Investigaciones Científicas (IVIC), Apartado 21827, Caracas, 1020-A, Venezuela

    Carlos Bello-Rodriguez, Olga Wittig, Dylana Diaz-Solano & Jose E. Cardier

  2. Facultad de Ciencias, Universidad Central de Venezuela, Caracas, 1080, Venezuela

    Carlos Bello-Rodriguez

  3. Laboratory of Cellular Physiology, Centre of Biophysics and Biochemistry, Instituto Venezolano de Investigaciones Científicas (IVIC), Caracas, 1020-A, Venezuela

    Pura Bolaños

  4. Unidad de Terapia Celular - Laboratorio de Patología Celular y Molecular, Centro de Medicina Experimental, Instituto Venezolano de Investigaciones Científicas (IVIC), Apartado 20632, Caracas, 1020-A, Venezuela

    Jose E. Cardier

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  1. Carlos Bello-Rodriguez
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  2. Olga Wittig
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Corresponding author

Correspondence to Jose E. Cardier.

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The authors declare that they have no conflict of interest.

Research involving human participants and/or animals

All animal experimentation was performed in accordance with institutional guidelines. MSCs used in this work were from healthy individuals treated for bone regeneration due to seudoarthrosis, secondary to a fracture, who authorized its use by signing informed consent.

Ethical approval

All applicable international, national and/or institutional guidelines for the care and use of animals were followed. This project was approved by the Bioethic Committee for Animal Research (Comisión de Bioética para Investigación en Animales, COBIANIM. No. 2014-15) of IVIC (Instituto Venezolano de Investigaciones Cientificas).

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Bello-Rodriguez, C., Wittig, O., Diaz-Solano, D. et al. A 3D construct based on mesenchymal stromal cells, collagen microspheres and plasma clot supports the survival, proliferation and differentiation of hematopoietic cells in vivo. Cell Tissue Res 382, 499–507 (2020). https://doi.org/10.1007/s00441-020-03265-y

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  • DOI: https://doi.org/10.1007/s00441-020-03265-y

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