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Tytuł:
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Combined approach for characterization and quality assessment of rabbit bone marrow-derived mesenchymal stem cells intended for gene banking.
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Autorzy:
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Vašíček J; NPPC - Research Institute for Animal Production in Nitra, Hlohovecká 2, 951 41 Lužianky, Slovak Republic; Research Centre AgroBioTech, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2, 949 76 Nitra, Slovak Republic; Faculty of Biotechnology and Food Science, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2, 949 76 Nitra, Slovak Republic. Electronic address: .
Kováč M; Faculty of Biotechnology and Food Science, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2, 949 76 Nitra, Slovak Republic.
Baláži A; NPPC - Research Institute for Animal Production in Nitra, Hlohovecká 2, 951 41 Lužianky, Slovak Republic.
Kulíková B; NPPC - Research Institute for Animal Production in Nitra, Hlohovecká 2, 951 41 Lužianky, Slovak Republic.
Tomková M; Faculty of Biotechnology and Food Science, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2, 949 76 Nitra, Slovak Republic.
Olexiková L; NPPC - Research Institute for Animal Production in Nitra, Hlohovecká 2, 951 41 Lužianky, Slovak Republic.
Čurlej J; Faculty of Biotechnology and Food Science, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2, 949 76 Nitra, Slovak Republic.
Bauer M; NPPC - Research Institute for Animal Production in Nitra, Hlohovecká 2, 951 41 Lužianky, Slovak Republic; Faculty of Natural Sciences, Constantine the Philosopher University in Nitra, Nábrežie mládeže 91, 949 74 Nitra, Slovak Republic.
Schnabl S; Department, of Internal Medicine I, Division of Haematology and Haemostaseology, Comprehensive Cancer Centre Vienna, Drug and Target Screening Unit DTSU, Medical University of Vienna, Waehringer Guertel 18-20, Vienna, A-1090, Austria.
Hilgarth M; Department, of Internal Medicine I, Division of Haematology and Haemostaseology, Comprehensive Cancer Centre Vienna, Drug and Target Screening Unit DTSU, Medical University of Vienna, Waehringer Guertel 18-20, Vienna, A-1090, Austria.
Hubmann R; Department, of Internal Medicine I, Division of Haematology and Haemostaseology, Comprehensive Cancer Centre Vienna, Drug and Target Screening Unit DTSU, Medical University of Vienna, Waehringer Guertel 18-20, Vienna, A-1090, Austria.
Shehata M; Department, of Internal Medicine I, Division of Haematology and Haemostaseology, Comprehensive Cancer Centre Vienna, Drug and Target Screening Unit DTSU, Medical University of Vienna, Waehringer Guertel 18-20, Vienna, A-1090, Austria.
Makarevich AV; NPPC - Research Institute for Animal Production in Nitra, Hlohovecká 2, 951 41 Lužianky, Slovak Republic.
Chrenek P; NPPC - Research Institute for Animal Production in Nitra, Hlohovecká 2, 951 41 Lužianky, Slovak Republic; Faculty of Biotechnology and Food Science, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2, 949 76 Nitra, Slovak Republic; Faculty of Animal Breeding and Biology, UTP University of Science and Technology, Mazowiecka 28, 85-084 Bydgoszcz, Poland.
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Źródło:
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New biotechnology [N Biotechnol] 2020 Jan 25; Vol. 54, pp. 1-12. Date of Electronic Publication: 2019 Aug 07.
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Typ publikacji:
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Journal Article
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Język:
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English
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Imprint Name(s):
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Original Publication: Amsterdam : Elsevier
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MeSH Terms:
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Biological Specimen Banks/*standards
Bone Marrow Cells/*cytology
Mesenchymal Stem Cells/*cytology
Rabbits/*genetics
Animals ; Antigens, CD/metabolism ; Bone Marrow Cells/metabolism ; Cryopreservation ; Mesenchymal Stem Cells/metabolism ; Phenotype ; RNA, Messenger/genetics ; Rabbits/classification ; Reverse Transcriptase Polymerase Chain Reaction
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Contributed Indexing:
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Keywords: Cryopreservation; Flow cytometry; MSCs; PCR; Rabbit; TEM
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Substance Nomenclature:
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0 (Antigens, CD)
0 (RNA, Messenger)
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Entry Date(s):
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Date Created: 20190811 Date Completed: 20200224 Latest Revision: 20200224
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Update Code:
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20240104
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DOI:
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10.1016/j.nbt.2019.08.001
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PMID:
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31400479
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Rabbit mesenchymal stem cells (rMSCs) are promising agents for the preservation of genetic biodiversity in domestic rabbit breeds. However, rMSCs must meet certain requirements to be used for cryopreservation in animal gene banks. Currently, there are numerous discrepancies in the published data regarding the rMSC phenotype, which may complicate efforts to evaluate their purity and suitability for reuse after cryopreservation in gene and tissue banks. We propose a combined approach (flow cytometry, PCR, differentiation and ultrastructure studies) for the characterization and recovery of rMSCs after cryopreservation. Flow cytometric analyses of rMSCs confirmed the expression of CD29, CD44, vimentin, desmin and α-SMA. RT-PCR revealed the expression of other markers at the mRNA level (SSEA-4, CD73, CD90, CD105, CD146 and CD166). rMSCs showed efficient multilineage differentiation into adipo-, chondro- and osteogenic lineages, SOX2 expression (pluripotency) and typical MSC morphology and ultrastructure. The confirmed rMSCs were subsequently used for cryopreservation. Efficient recovery of rMSCs after cryogenic freezing was demonstrated by high cell viability, normal ultrastructure of reseeded rMSCs, high expression of CD29 and CD44 and lineage differentiation capacity. The proposed combined approach could be used for characterization, cryopreservation and recovery of rMSCs as genetic resources for native rabbit breeds.
(Copyright © 2019. Published by Elsevier B.V.)
