Cryopreservation of heat-shocked canine adipose-derived mesenchymal stromal cells with 10% dimethyl sulfoxide and 40% serum results in better viability,proliferation, anti-oxidation,and in-vitro differentiation |
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| Institution: | 1. Department of Pharmacology, Pennsylvania State University, College of Medicine, Hershey, PA, USA;2. Department of Pathology, Pennsylvania State University, College of Medicine, Hershey, PA, USA;3. Department of Comparative Medicine, Pennsylvania State University, College of Medicine, Hershey, PA, USA;4. Division of Pulmonary and Critical Care Medicine, Pennsylvania State University, College of Medicine, Hershey, PA, USA;1. Institute of Beijing Animal Science and Veterinary, Chinese Academy of Agricultural Science, Beijing, 100193, China;2. Department of Life Science, Department of laboratory medicine, Bengbu Medical College, Bengbu, 233030, China;3. Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, 150001, China;1. Área Inmunología, Facultad de Veterinaria, Universidad de la República, 11600, Montevideo, Uruguay;2. Laboratorio de Genética Bioquímica (LAGENBIO), Instituto Agroalimentario de Aragón IA2 - Instituto de Investigación Sanitaria de Aragón IIS, Universidad de Zaragoza, 50013, Zaragoza, Spain;3. Servicio de Cirugía y Medicina Equina, Hospital Veterinario, Universidad de Zaragoza, C/Miguel Servet, 177, 50013, Zaragoza, Spain;1. Universidad de Concepción, Campus Chillan, Faculty of Veterinary Sciences, Department of Animal Science, Laboratorio de Biotecnologia Animal, Chile;2. Universidad de Concepción, Campus Chillan, Faculty of Veterinary Sciences, Department of Animal Pathology, Chile;3. Universidad de Concepción, Campus Chillan, Faculty of Veterinary Sciences, Department of Clinical Sciences, Hospital de Animales Mayores, Chile;1. Research Institute for Animal Production in Nitra, NAFC, Hlohovecká 2, 951 41 Lu?ianky, Slovak Republic;2. Faculty of Biotechnology and Food Science, Slovak University of Agriculture, Tr. A. Hlinku 2, 949 76 Nitra, Slovak Republic;3. Faculty of Natural Sciences, Constantine the Philosopher University in Nitra, Nábre?ie mláde?e 91, 949 74 Nitra, Slovak Republic;4. Faculty of Animal Breeding and Biology, University of Technology and Life Sciences, Al. prof. S. Kaliskiego 7, 85 796 Bydgoszcz, Poland |
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| Abstract: | Cryopreserved canine adipose-derived mesenchymal stromal cells (Ad-MSCs) can be used instantly in dogs for clinical uses. However, cryopreservation results in a reduction of the cellular viability, proliferation, and anti-oxidation of post-thawed Ad-MSCs. Therefore, there is a need for in-vitro procedure to improve post-thawed Ad-MSCs’ viability, proliferation, anti-oxidation, and differentiation capacity. In this study, fresh-Ad-MSCs were activated with heat shock, hypoxia (5% O2), or hypoxia (5% O2) + heat shock treatments. The results showed that compared to the other treatments, heat shock significantly improved the proliferation rate, anti-oxidation, heat shock proteins and growth factors expressions of canine-fresh-Ad-MSCs. Consequently, fresh-Ad-MSCs were heat-shocked and then cryopreserved with different combinations of dimethyl sulfoxide (Me2SO) and fetal bovine serum (FBS) to determine the combination that could effectively preserve the cellular viability, proliferation, anti-oxidation and differentiation capacity of Ad-MSCs after cryopreservation. We found that C-HST-Ad-MSCs cryopreserved with 10% Me2SO + 40% FBS presented significantly (p < 0.05) improved cellular viability, proliferation rate, anti-oxidant capacity, and differentiation potential as compared to C-HST-Ad-MSCs cryopreserved with 1% Me2SO + 10% FBS or 1% Me2SO alone or control. We concluded, heat shock treatment is much better to enhance the characteristics of fresh-Ad-MSCs than other treatments, moreover, C-HST-Ad-MSCs in 10% Me2SO + 40% FBS showed better results compared to other cryopreserved groups. However, future work is required to optimize the expression of heat shock proteins, which would further improve the characteristics of fresh- and cryopreserved-HST-Ad-MSCs and reduce the dependency on Me2SO and FBS. |
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| Keywords: | Mesenchymal stromal cells Heat shock treatment Hypoxia Cryopreservation Anti-oxidation In vitro differentiation MSCs"} {"#name":"keyword" "$":{"id":"kwrd0045"} "$$":[{"#name":"text" "_":"Mesenchymal stem cells Ad-MSCs"} {"#name":"keyword" "$":{"id":"kwrd0055"} "$$":[{"#name":"text" "_":"Adipose-derived mesenchymal stromal cells HSP"} {"#name":"keyword" "$":{"id":"kwrd0065"} "$$":[{"#name":"text" "_":"Heat shock protein HST-Ad-MSCs"} {"#name":"keyword" "$":{"id":"kwrd0075"} "$$":[{"#name":"text" "_":"Heat shock treated-Adipose-derived mesenchymal stromal cells F-HST-Ad-MSCs"} {"#name":"keyword" "$":{"id":"kwrd0085"} "$$":[{"#name":"text" "_":"Fresh-Heat shock treated-Adipose-derived mesenchymal stromal cells C-HST-Ad-MSCs"} {"#name":"keyword" "$":{"id":"kwrd0095"} "$$":[{"#name":"text" "_":"Cryopreserved-Heat shock treated-Adipose-derived mesenchymal stromal cells HT-Ad-MSCs"} {"#name":"keyword" "$":{"id":"kwrd0105"} "$$":[{"#name":"text" "_":"Hypoxia-treated-Adipose-derived mesenchymal stromal cells HT + HST-Ad-MSCs"} {"#name":"keyword" "$":{"id":"kwrd0115"} "$$":[{"#name":"text" "_":"Hypoxia treatment and heat shock treated-Adipose-derived mesenchymal stromal cells |
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