Cryopreservation based on freezing protocols for the long-term storage of microencapsulated myoblasts |
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| Authors: | Ainhoa Murua Gorka Orive Rosa Ma. Hernández José Luis Pedraz |
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| Affiliation: | 1. Laboratory of Pharmacy and Pharmaceutical Technology, Faculty of Pharmacy, University of the Basque Country, 01006 Vitoria-Gasteiz, Spain;2. Networking Biomedical Research Center on Bioengineering, Biomaterials and Nanomedicine, CIBER-BBN, SLFPB-EHU, 01006 Vitoria-Gasteiz, Spain;1. Department of Surgery, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA;2. Department of Urology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA;3. Department of Obstetrics and Gynecology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA;4. Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA;5. Shriners Hospital for Children, Boston, MA 02114, USA;1. Department of Biomedical Engineering, The Ohio State University, Columbus, OH 43210, United States;2. Davis Heart and Lung Research Institute, The Ohio State University, Columbus, OH 43210, United States;3. Department of Mechanical and Aerospace Engineering, The Ohio State University, Columbus, OH 43210, United States;4. Comprehensive Cancer Center, The Ohio State University, Columbus, OH 43210, United States;5. Center for Biomedical Engineering, University of Science and Technology of China, Hefei, Anhui 230027, China |
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| Abstract: | One important challenge in biomedicine is the ability to cryogenically preserve not only cells, but also tissue-engineered constructs. In the present paper, alginate-poly-l-lysine-alginate (APA) microcapsules containing erythropoietin (Epo)-secreting C2C12 myoblasts were elaborated, characterized and tested both in vitro and in vivo. Dimethylsulfoxide (DMSO) was selected as cryoprotectant to evaluate the maintenance of physiological activity of cryopreserved microencapsulated myoblasts employing procedures based on freezing protocols up to a 45-day cryopreservation period. High chemical resistance of the cryopreserved microcapsules was observed using 10% DMSO as cryoprotectant following a standard slow-cooling procedure. Although a 42% reduction in Epo release from the microencapsulated cells was observed in comparison with the non-cryopreserved group, the in vivo biocompatibility and functionality of the encapsulated cells subcutaneously implanted in Balb/c mice was corroborated by high and sustained hematocrit levels over 194 days and lacking immunosuppressive protocols. No major host reaction was observed. Based on the results obtained in our study, a slow-cooling protocol using 10% DMSO as cryoprotectant (confirmed for cryopreservation periods up to 45 days) might be considered a suitable therapeutic strategy if the long-term storage of microencapsulated cells, such as C2C12 myoblasts is pretended. |
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