title: Expanded cryopreserved mesenchymal stromal cells as an optimal source for graft-versus-host disease treatment.
abstract: Mesenchymal stromal cells (MSC) are fibroblast-like cells present in different types of tissues. Their immunomodulatory potential represents a promising method for post-transplant immunotherapy in the treatment of GVHD (graft-versus-host disease) with suboptimal response to standard immunosuppression. In this study we tested influence of 1-8 month-long cryopreservation on ability of MSC to suppress activation of non-specifically stimulated lymphocytes. We did not observe any changes in proliferation capacity of MSC after thawing. Lymphocytes metabolic activity was inhibited by 30% and number of dividing cells was three times smaller in the presence of MSC. Two activation markers were studied (CD25 and CD69) to confirm preservation of functional cell integrity. Expression of CD25 antigen on CD3(+)CD4(+) and CD3(+)CD4(-) cells was decreased in all co-cultivated samples. Level of CD69 expression on CD3(+)CD4(+) cells was lower in samples with added MSC (10-15% on day +2) but without reaching statistical significance. The lower expression (approximately 5%) was observed also on CD4-cells. The study confirms the preservation of immunomodulatory properties of cryopreserved and re-expanded MSC. Aliquots with cryopreserved cells can represent an optimal source for a quick preparation of MSC cell product with the possibility to apply the same cells repeatedly.
authors.affiliation: Department of Hematology and Oncology, Charles University in Prague, Medical School and Teaching Hospital in Pilsen, Pilsen, Czech Republic; Biomedical Centre, Faculty of Medicine in Pilsen, Charles University in Prague, Pilsen, Czech Republic. Electronic address: holubovam@fnplzen.cz.
authors.forename: Monika
authors.initials: M
authors.lastname: Holubova
authors.affiliation: Department of Hematology and Oncology, Charles University in Prague, Medical School and Teaching Hospital in Pilsen, Pilsen, Czech Republic; Biomedical Centre, Faculty of Medicine in Pilsen, Charles University in Prague, Pilsen, Czech Republic.
authors.forename: Daniel
authors.initials: D
authors.lastname: Lysak
authors.affiliation: Institute of Immunology and Allergology, Charles University in Prague, Medical School and Teaching Hospital in Pilsen, Pilsen, Czech Republic.
authors.forename: Tomas
authors.initials: T
authors.lastname: Vlas
authors.affiliation: Laboratory of Immunotherapy, Institute of Microbiology, Academy of Sciences of the Czech Republic, v.v.i, Prague, Czech Republic.
authors.forename: Luca
authors.initials: L
authors.lastname: Vannucci
authors.affiliation: Czech National Marrow Donor Registry (CS-2), Pilsen, Czech Republic.
authors.forename: Pavel
authors.initials: P
authors.lastname: Jindra
mesh_terms: D049109:Cell Proliferation; D015925:Cryopreservation; D006086:Graft vs Host Disease / Q000628:therapy*; D006801:Humans; D016130:Immunophenotyping; D008213:Lymphocyte Activation; D059630:Mesenchymal Stem Cells / Q000166:cytology*
keywords: Cryopreservation; Immunomodulation; Mesenchymal stromal cells