Decoding immunomodulatory interactions: Exploring the crosstalk between bone marrow-derived mesenchymal steam cells (BM-MSCs) out the bag and peripheral blood mononuclear cells (PBMCs) for potential in-vivo immunosuppresion

Monteiro, Raquel Leão

http://hdl.handle.net/10400.22/29806

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Authors

Monteiro, Raquel Leão

Advisor(s)

Lopes, Sérgio Manuel Bernardo Machado

Vieira, Ana Filipa Quintela

Abstract(s)

Allogenic hematopoietic stem cell transplant (Allo-HSCT) is crucial for hematological diseases, yet chronic complications may arise as donor imune cells start attacking patient tissues. A lab barrier to studying this alloreactivity is the absence of the hematopoietic stem cell (HSC) niche. Here, we employed boné marrow-derived mesenchymal stem cells (BM-MSCs) from discarded collection bags to assess their potential in modulating peripheral blood mononuclear cell (PBMC) proliferation in-vitro and decode occuring interactions to propose mechanisms of in-vivo immunomodulation. Responder PBMCs, labelled with CFSE dye, were co-cultured for 6 days with irradiated stimulator cell in a 2:1 ratio to mimic alloreactivity, with or without BM-MSCs. Flow cytometry tracked BM-MSCs phenotype and PBMC proliferation/ surface marker expression, while resazurin assays monitored metabolic activity over time. CFSE immunofluorescence and phase-contrast images recorded PBMC proliferation interactions. BM-MSCs expanded successfully, maintaining adequate phenotype. Co-cultures exhibited fewer CFSE-stained PBMCs, equating to na 18% reduction in responder cells, with enhanced metabolic activity. Immunophenotyping revealed downregulation of T cell subsets and increased PD-1 expression, suggesting immunosuppressive effects. These findings underscore BM-MSCs’ immunomodulatory potential, offering a celular therapeutic avenue. Repurposing collection bags as a source of BM-MSCs also offers a practical solution, reducing medical waste and enhancing cost efficiency.