title: Cryopreservation of microencapsulated murine mesenchymal stem cells genetically engineered to secrete erythropoietin.
abstract: The ability to cryopreserve and store for long term the structure and function of therapeutic cells and tissues plays a pivotal role in clinical medicine. In fact, it is an essential pre-requisite for the commercial and clinical application of stem cells since preserves cells at low temperature and creates a reserve for future uses. This requisite may also affect the encapsulated stem cells. Several parameters should be considered on encapsulated cell cryopreservation such as the time and temperature during the cryopreservation process, or the cryoprotectant solutions used. In this study, we have compared the influence of penetrating and nonpenetrating cryoprotectants on the viability and functionality of encapsulated mesenchymal stem cells genetically modified to secrete erythropoeitin. Several cryoprotectant solutions combining DMSO, glycerol and trehalose at different concentrations were studied. Although almost no differences among the studied cryoprotectant solutions were observed on the differentiation potential of encapsulated mesenchymal stem cells, the penetrating cryoprotectant DMSO at a concentration of 10% displayed the best viability and erythropoietin secretion profile compared to the other cryoprotectant solutions. These results were confirmed after subcutaneous implantation of thawed encapsulated mesenchymal stem cells secreting erythropoeitin on Balb/c mice. The hematocrit levels of these animals increased to similar levels of those detected on animals transplanted with noncryopreserved encapsulated cells. Therefore, DMSO 10% represents the most suitable cryoprotectant solution among the solutions here studied, for encapsulated mesenchymal stem cells cryopreservation and its translation into the clinic. Similar studies should be performed for the encapsulation of other cell types before they can be translated into the clinic.
authors.affiliation: NanoBioCel Group, Laboratory of Pharmacy and Pharmaceutical Technology, Faculty of Pharmacy, University of the Basque Country, UPV/EHU, Vitoria-Gasteiz, Spain; Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine, CIBER-BBN, Vitoria-Gasteiz, Spain. Electronic address: haritz.g.i@gmail.com.
authors.forename: Haritz
authors.initials: H
authors.lastname: Gurruchaga
authors.affiliation: NanoBioCel Group, Laboratory of Pharmacy and Pharmaceutical Technology, Faculty of Pharmacy, University of the Basque Country, UPV/EHU, Vitoria-Gasteiz, Spain; Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine, CIBER-BBN, Vitoria-Gasteiz, Spain. Electronic address: jeciriza@gmail.com.
authors.forename: Jesús
authors.initials: J
authors.lastname: Ciriza
authors.affiliation: NanoBioCel Group, Laboratory of Pharmacy and Pharmaceutical Technology, Faculty of Pharmacy, University of the Basque Country, UPV/EHU, Vitoria-Gasteiz, Spain; Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine, CIBER-BBN, Vitoria-Gasteiz, Spain. Electronic address: laura.saenzdelburgo@ehu.es.
authors.forename: Laura
authors.initials: L
authors.lastname: Saenz Del Burgo
authors.affiliation: Laboratory of Cell Therapy, Division of Oncology, Foundation for Applied Medical Research, University of Navarra, Pamplona 31008, Spain. Electronic address: jrrodriguez@unav.es.
authors.forename: Juan Roberto
authors.initials: JR
authors.lastname: Rodriguez-Madoz
authors.affiliation: NanoBioCel Group, Laboratory of Pharmacy and Pharmaceutical Technology, Faculty of Pharmacy, University of the Basque Country, UPV/EHU, Vitoria-Gasteiz, Spain; Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine, CIBER-BBN, Vitoria-Gasteiz, Spain. Electronic address: edorta.santos@ehu.es.
authors.forename: Edorta
authors.initials: E
authors.lastname: Santos
authors.affiliation: Hematology and Cell Therapy, Clinica Universidad de Navarra, University of Navarra, Pamplona 31008, Spain. Electronic address: fprosper@unav.es.
authors.forename: Felipe
authors.initials: F
authors.lastname: Prosper
authors.affiliation: NanoBioCel Group, Laboratory of Pharmacy and Pharmaceutical Technology, Faculty of Pharmacy, University of the Basque Country, UPV/EHU, Vitoria-Gasteiz, Spain; Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine, CIBER-BBN, Vitoria-Gasteiz, Spain. Electronic address: rosa.hernandez@ehu.es.
authors.forename: Rosa María
authors.initials: RM
authors.lastname: Hernández
authors.affiliation: NanoBioCel Group, Laboratory of Pharmacy and Pharmaceutical Technology, Faculty of Pharmacy, University of the Basque Country, UPV/EHU, Vitoria-Gasteiz, Spain; Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine, CIBER-BBN, Vitoria-Gasteiz, Spain. Electronic address: gorka.orive@ehu.es.
authors.forename: Gorka
authors.initials: G
authors.lastname: Orive
authors.affiliation: NanoBioCel Group, Laboratory of Pharmacy and Pharmaceutical Technology, Faculty of Pharmacy, University of the Basque Country, UPV/EHU, Vitoria-Gasteiz, Spain; Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine, CIBER-BBN, Vitoria-Gasteiz, Spain. Electronic address: joseluis.pedraz@ehu.es.
authors.forename: Jose Luis
authors.initials: JL
authors.lastname: Pedraz
mesh_terms: D000818:Animals; D048430:Cell Shape / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D015925:Cryopreservation; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004921:Erythropoietin / Q000235:genetics / Q000378:metabolism*; D005990:Glycerol / Q000494:pharmacology; D006400:Hematocrit; D006410:Hematopoiesis; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000187:drug effects* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D013997:Time Factors; D014162:Transfection; D014199:Trehalose / Q000494:pharmacology
chemical_list: D003451:Cryoprotective Agents; D004921:Erythropoietin; D014199:Trehalose; D005990:Glycerol; D004121:Dimethyl Sulfoxide
keywords: Cryopreservation; Mesenchymal stem cells; Microencapsulation; Nanotechnology