title: Diverse impact of xeno-free conditions on biological and regenerative properties of hUC-MSCs and their extracellular vesicles.
abstract: Growing evidence indicates that intracellular signaling mediated by extracellular vesicles (EVs) released by stem cells plays a considerable role in triggering the regenerative program upon transplantation. EVs from umbilical cord mesenchymal stem cells (UC-MSC-EVs) have been shown to enhance tissue repair in animal models. However, translating such results into clinical practice requires optimized EV collection procedures devoid of animal-originating agents. Thus, in this study, we analyzed the influence of xeno-free expansion media on biological properties of UC-MSCs and UC-MSC-EVs for future applications in cardiac repair in humans. Our results show that proliferation, differentiation, phenotype stability, and cytokine secretion by UC-MSCs vary depending on the type of xeno-free media. Importantly, we found distinct molecular and functional properties of xeno-free UC-MSC-EVs including enhanced cardiomyogenic and angiogenic potential impacting on target cells, which may be explained by elevated concentration of several pro-cardiogenic and pro-angiogenic microRNA (miRNAs) present in the EVs. Our data also suggest predominantly low immunogenic capacity of certain xeno-free UC-MSC-EVs reflected by their inhibitory effect on proliferation of immune cells in vitro. Summarizing, conscious selection of cell culture conditions is required to harvest UC-MSC-EVs with the optimal desired properties including enhanced cardiac and angiogenic capacity, suitable for tissue regeneration.
  
  
  
  Type of xeno-free media influences biological properties of UC-MSCs in vitro. Certain xeno-free media promote proliferation and differentiation ability of UC-MSCs. EVs collected from xeno-free cultures of UC-MSCs are biologically active. Xeno-free UC-MSC-EVs enhance cardiac and angiogenic potential of target cells. Type of xeno-free media determines immunomodulatory effects mediated by UC-MSC-EVs.
authors.affiliation: Department of Cell Biology, Faculty of Biochemistry, Biophysics, and Biotechnology, Jagiellonian University, 30-387, Krakow, Poland. sylwia.bobis@uj.edu.pl.
authors.forename: Sylwia
authors.identifier: 0000-0002-2200-6767
authors.initials: S
authors.lastname: Bobis-Wozowicz
authors.affiliation: Department of Cell Biology, Faculty of Biochemistry, Biophysics, and Biotechnology, Jagiellonian University, 30-387, Krakow, Poland.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Kmiotek
authors.affiliation: Department of Cell Biology, Faculty of Biochemistry, Biophysics, and Biotechnology, Jagiellonian University, 30-387, Krakow, Poland.
authors.forename: Karolina
authors.initials: K
authors.lastname: Kania
authors.affiliation: Department of Cell Biology, Faculty of Biochemistry, Biophysics, and Biotechnology, Jagiellonian University, 30-387, Krakow, Poland.|Malopolska Centre of Biotechnology, 30-387, Krakow, Poland.
authors.forename: Elzbieta
authors.initials: E
authors.lastname: Karnas
authors.affiliation: Department of Cell Biology, Faculty of Biochemistry, Biophysics, and Biotechnology, Jagiellonian University, 30-387, Krakow, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Labedz-Maslowska
authors.affiliation: Malopolska Centre of Biotechnology, 30-387, Krakow, Poland.
authors.forename: Malgorzata
authors.initials: M
authors.lastname: Sekula
authors.affiliation: Department of Physical Biochemistry, Faculty of Biochemistry, Biophysics, and Biotechnology, Jagiellonian University, 30-387, Krakow, Poland.
authors.forename: Sylwia
authors.initials: S
authors.lastname: Kedracka-Krok
authors.affiliation: Department of Pediatric Cardiac Surgery, Polish-American Children's Hospital, 30-663, Krakow, Poland.
authors.forename: Jacek
authors.initials: J
authors.lastname: Kolcz
authors.affiliation: Polish Stem Cell Bank, 00-131, Warsaw, Poland.
authors.forename: Dariusz
authors.initials: D
authors.lastname: Boruczkowski
authors.affiliation: Department of Cell Biology, Faculty of Biochemistry, Biophysics, and Biotechnology, Jagiellonian University, 30-387, Krakow, Poland.
authors.forename: Zbigniew
authors.initials: Z
authors.lastname: Madeja
authors.affiliation: Department of Cell Biology, Faculty of Biochemistry, Biophysics, and Biotechnology, Jagiellonian University, 30-387, Krakow, Poland. ewa.zuba-surma@uj.edu.pl.
authors.forename: Ewa K
authors.initials: EK
authors.lastname: Zuba-Surma
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free / Q000737:chemistry / Q000494:pharmacology*; D016207:Cytokines / Q000378:metabolism; D000067128:Extracellular Vesicles / Q000187:drug effects* / Q000502:physiology; D006321:Heart / Q000502:physiology*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000187:drug effects* / Q000502:physiology; D035683:MicroRNAs / Q000235:genetics; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D012038:Regeneration; D014470:Umbilical Cord / Q000166:cytology*
chemical_list: D016895:Culture Media, Serum-Free; D016207:Cytokines; D035683:MicroRNAs; D000255:Adenosine Triphosphate
keywords: Extracellular vesicles; Heart regeneration; Immunomodulation; Mesenchymal stem cells; Umbilical cord; Xeno-free