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Histochemistry and Cell Biology

, Volume 146, Issue 2, pp 153–165 | Cite as

Mesenchymal stromal cells support the viability and differentiation of thymocytes through direct contact in autologous co-cultures

  • Seyed Mohammad Reza Azghadi
  • Maria Suciu
  • Alexandra Teodora GruiaEmail author
  • Lucian Barbu-Tudoran
  • Mirabela Iustina Cristea
  • Ani Aurora Mic
  • Danina Muntean
  • Dragos Vasile Nica
  • Felix Aurel Mic
Original Paper

Abstract

The development of thymocytes and generation of mature T cells is a complex process that requires spatio-temporal interactions of thymocytes with the other cells of the thymus microenvironment. Recently, mesenchymal stromal cells were isolated from the neonatal human thymus and differentiated into chondrogenic, osteogenic, and adipogenic lineages, just like their bone marrow counterparts. However, their function in thymocyte homeostasis is unknown. In our autologous co-cultures of rat mesenchymal stromal cells and thymocytes, the stromal cells preserve the viability of cultured thymocytes and stimulate the development of CD4−CD8− double-negative and the maturation of mainly CD4+ single-positive thymocytes. Thymocytes also influence the stemness of bone marrow mesenchymal stromal cells, as their expression of CD44, a marker associated with cellular proliferation and migration, is reduced in co-cultures. Mesenchymal stromal cells’ influence on thymocyte development requires direct physical contact between the two cells and is not mediated by a soluble factor. When the two types of cells were physically separated, the stimulative effects of mesenchymal stromal cells on thymocytes did not occur. Electron microscopy confirmed the close contact between the membranes of thymocytes and mesenchymal stromal cells. Our experiments suggest that membrane exchanges could occur between mesenchymal stromal cells and thymocytes, such as the transfer of CD44 from mesenchymal stromal cells to the thymocytes, but its functional significance for thymocytes development remains to be established. These results suggest that mesenchymal stromal cells could normally be a part of the in vivo thymic microenvironment and form a niche that could sustain and guide the development of thymocytes.

Keywords

Mesenchymal stromal cells Co-cultures Maturation Thymocytes Viability 

Abbreviations

DMEM

Dulbecco’s modified Eagle’s medium

FCS

Fetal calf serum

PBS

Phosphate-buffered saline

BMSCs

Bone marrow-derived mesenchymal stromal cells

DN

CD4−CD8− double-negative thymocytes

DP

CD4+CD8+ double-positive thymocytes

CD4+ SP

CD4+ single-positive thymocytes

CD8+ SP

CD8+ single-positive thymocytes

TCR

T cell receptor

FSC

Forward scatter

SSC

Side scatter

qPCR

Real-time (quantitative) polymerase chain reaction

TEM

Transmission electron microscopy

SEM

Scanning electron microscopy

Notes

Acknowledgments

This work was supported by a Grant of the Romanian National Authority for Scientific Research, CNCS—UEFISCDI, Project Number PN-II-ID-PCE-2011-3-0571, awarded to FAM. DVN was co-financed from the European Social Fund through Sectorial Operational Programme Human Resources Development 2007–2013, project number POSDRU/CPP107/DMI 1.5/S/77082, “Doctoral Scholarships for eco-economy and bio-economic complex training to ensure the food and feed safety and security of anthropogenic ecosystems.” This paper is partly supported by the Sectorial Operational Programme Human Resources Development (SOPHRD), financed by the European Social Fund and the Romanian Government under contract number POSDRU 141531. MS was supported by the strategic grant POSDRU/159/1.5/S/133391, Project “Doctoral and Post-doctoral programs of excellence for highly qualified human resources training for research in the field of Life sciences, Environment and Earth Science” co-financed by the European Social Fund within the Sectorial Operational Program Human Resources Development 2007–2013.

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Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Seyed Mohammad Reza Azghadi
    • 1
  • Maria Suciu
    • 2
    • 3
  • Alexandra Teodora Gruia
    • 4
    Email author
  • Lucian Barbu-Tudoran
    • 5
  • Mirabela Iustina Cristea
    • 4
  • Ani Aurora Mic
    • 1
    • 6
  • Danina Muntean
    • 7
  • Dragos Vasile Nica
    • 4
    • 8
  • Felix Aurel Mic
    • 1
  1. 1.Department of Functional SciencesUniversity of Medicine and Pharmacy “Victor Babes” TimisoaraTimisoaraRomania
  2. 2.Electron Microscopy Integrated LaboratoryNational Institute for Research and Development of Isotopic and Molecular TechnologiesCluj-NapocaRomania
  3. 3.Molecular Biology and Biotechnology Department, Faculty of Biology and GeologyBabeş-Bolyai UniversityCluj-NapocaRomania
  4. 4.Regional Center of Immunology of TransplantationEmergency Clinical County Hospital TimisoaraTimisoaraRomania
  5. 5.Center of Electron Microscopy, Faculty of Biology and GeologyBabes-Bolyai UniversityCluj-NapocaRomania
  6. 6.INCD “Victor Babes”BucharestRomania
  7. 7.Department of Pathophysiology, Center for Translational Research and Systems Medicine“Victor Babes” University of Medicine and PharmacyTimisoaraRomania
  8. 8.Faculty of Animal Sciences and BiotechnologiesBanat’s University of Agricultural Sciences and Veterinary MedicineTimisoaraRomania

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