title: Effects of Freeze-Thawing and Intravenous Infusion on Mesenchymal Stromal Cell Gene Expression.
abstract: Mesenchymal stromal cells (MSC) are increasingly used as an investigative therapeutic product for immune disorders and degenerative disease. Typically, MSC are isolated from human tissue, expanded in culture, and cryopreserved until usage. The safety and efficacy of MSC therapy will depend on the phenotypical and functional characteristics of MSC. The freeze-thawing procedure may change these characteristics. Furthermore, the cells encounter a microenvironment after administration that may impact their properties. It has been demonstrated that the majority of MSC localize to the lungs after intravenous infusion, making this the site to study the effects of the in vivo milieu on administered MSC. In this study, we investigated the effect of freeze-thawing and the mouse lung microenvironment on human adipose tissue-derived MSC. There were effects of freeze-thawing on the whole genome expression profile of MSC, although the effects did not exceed interdonor differences. There were no major changes in the expression of hemostatic regulators on transcriptional level, but significantly increased expression of procoagulant tissue factor on the surface of thawed adipose MSC, correlating with increased procoagulant activity of thawed cells. Exposure for 2 h to the lung microenvironment had a major effect on MSC gene expression and affected several immunological pathways. This indicates that MSC undergo functional changes shortly after infusion and this may influence the efficacy of MSC to modulate inflammatory responses. The results of this study demonstrate that MSC rapidly alter in response to the local milieu and disease-specific conditions may shape MSC after administration.
authors.affiliation: 1 Nephrology and Transplantation, Department of Internal Medicine, Erasmus Medical Center , Rotterdam, the Netherlands .
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Hoogduijn
authors.affiliation: 1 Nephrology and Transplantation, Department of Internal Medicine, Erasmus Medical Center , Rotterdam, the Netherlands .
authors.forename: Samantha F H
authors.initials: SF
authors.lastname: de Witte
authors.affiliation: 1 Nephrology and Transplantation, Department of Internal Medicine, Erasmus Medical Center , Rotterdam, the Netherlands .
authors.forename: Franka
authors.initials: F
authors.lastname: Luk
authors.affiliation: 2 Center for Biomics, Erasmus Medical Center , Rotterdam, the Netherlands .
authors.forename: Mirjam C G N
authors.initials: MC
authors.lastname: van den Hout-van Vroonhoven
authors.affiliation: 3 Department of Dermatology and Venerology, Lund University , Stockholm, Sweden .
authors.forename: Lech
authors.initials: L
authors.lastname: Ignatowicz
authors.affiliation: 4 Department of Nephrology and Intensive Care Medicine, Charité Universtätsmedizin Berlin , Berlin, Germany .
authors.forename: Rusan
authors.initials: R
authors.lastname: Catar
authors.affiliation: 1 Nephrology and Transplantation, Department of Internal Medicine, Erasmus Medical Center , Rotterdam, the Netherlands .
authors.forename: Tanja
authors.initials: T
authors.lastname: Strini
authors.affiliation: 1 Nephrology and Transplantation, Department of Internal Medicine, Erasmus Medical Center , Rotterdam, the Netherlands .
authors.forename: Sander S
authors.initials: SS
authors.lastname: Korevaar
authors.affiliation: 2 Center for Biomics, Erasmus Medical Center , Rotterdam, the Netherlands .
authors.forename: Wilfred F J
authors.initials: WF
authors.lastname: van IJcken
authors.affiliation: 1 Nephrology and Transplantation, Department of Internal Medicine, Erasmus Medical Center , Rotterdam, the Netherlands .
authors.forename: Michiel G H
authors.initials: MG
authors.lastname: Betjes
authors.affiliation: 1 Nephrology and Transplantation, Department of Internal Medicine, Erasmus Medical Center , Rotterdam, the Netherlands .
authors.forename: Marcella
authors.initials: M
authors.lastname: Franquesa
authors.affiliation: 5 Berlin-Brandenburg Center/School for Regenerative Therapies (BCRT/BSRT), Charité Universtätsmedizin Berlin , Berlin, Germany .|6 Division of Therapeutic Immunology (TIM), Department of Laboratory Medicine (LABMED), Karolinska Institutet , Stockholm, Sweden .
authors.forename: Guido
authors.initials: G
authors.lastname: Moll
authors.affiliation: 1 Nephrology and Transplantation, Department of Internal Medicine, Erasmus Medical Center , Rotterdam, the Netherlands .
authors.forename: Carla C
authors.initials: CC
authors.lastname: Baan
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D018925:Chemokines / Q000235:genetics / Q000378:metabolism; D015925:Cryopreservation; D005260:Female; D005615:Freezing; D006706:Homeostasis; D006801:Humans; D007262:Infusions, Intravenous; D008168:Lung / Q000276:immunology / Q000378:metabolism; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells; D053858:Metabolic Networks and Pathways; D008810:Mice, Inbred C57BL; D059467:Transcriptome
chemical_list: D018925:Chemokines