title: Closed Vitrification System as A Platform for Cryopreservation of Tissue Engineered Constructs.
abstract: BACKGROUND
  Cryopreservation of mesenchymal stromal cells (MSCs) and MSCs-based tissue engineered constructs (TECs) is a promising strategy for regenerative medicine.
  
  
  OBJECTIVE
  To examine vitrification system consisting of multicomponent vitreous solution, closed type container, human adult MSCs and two-step exposure procedure as a platform for cryopreservation of MSCs-based TECs.
  
  
  METHODS
  Vitrification properties of solutions were studied by visual analysis and calorimetry. Viability (trypan blue, MTT-test), metabolic activity (Alamar Blue assay) and adhesion of cells were assessed both after exposure with vitreous solutions and following rapid cooling-thawing in standard cryovials.
  
  
  RESULTS
  The feasibility of the vitrification system was tested on MSCs suspensions (S-MSCs) and alginate encapsulated MSCs (AE-MSCs). The minimal concentrations of cryoprotectants, which allowed avoiding ice formation during rapid cooling and rewarming comprised 10 % for dimethylsulfoxide, 20 % for ethylene glycol, 20 % for 1.2-propanediol and 0.5 M sucrose. To achieve viability and metabolic activity rates of AE-MSCs comparable to S-MSCs after vitrification the extension of the exposure time within the same vitreous solution was sufficient. After vitrification both S-MSCs and AE-MSCs retained the capacity to osteogenic and adipogenic differentiation.
  
  
  CONCLUSIONS
  Data demonstrate that this vitrification system can be used as a platform for development of effective protocols for cryopreservation of MSCs-based TECs.
authors.affiliation: Institute for Problems of Cryobiology and Cryomedicine, NAS of Ukraine, Kharkov, Ukraine. n.a.trufan@gmail.com.
authors.forename: N A
authors.initials: NA
authors.lastname: Trufanova
authors.affiliation: Institute for Problems of Cryobiology and Cryomedicine, NAS of Ukraine, Kharkov, Ukraine.
authors.forename: V S
authors.initials: VS
authors.lastname: Zaikov
authors.affiliation: Institute for Problems of Cryobiology and Cryomedicine, NAS of Ukraine, Kharkov, Ukraine.
authors.forename: A V
authors.initials: AV
authors.lastname: Zinchenko
authors.affiliation: Institute for Problems of Cryobiology and Cryomedicine, NAS of Ukraine, Kharkov, Ukraine.
authors.forename: A Yu
authors.initials: AY
authors.lastname: Petrenko
authors.affiliation: Institute for Problems of Cryobiology and Cryomedicine, NAS of Ukraine, Kharkov, Ukraine. Department of Neuroscience, Institute of Experimental Medicine, Prague, Czech Republic.
authors.forename: Y A
authors.initials: YA
authors.lastname: Petrenko
mesh_terms: D002454:Cell Differentiation / Q000187:drug effects; D003451:Cryoprotective Agents; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects; D023822:Tissue Engineering / Q000295:instrumentation / Q000379:methods*; D058989:Vitrification
chemical_list: D003451:Cryoprotective Agents