Combined approach for characterization and quality assessment of rabbit bone marrow-derived mesenchymal stem cells intended for gene banking
Autor: | Susanne Schnabl, Miroslav Bauer, Martin Hilgarth, Maria Tomkova, Andrej Baláži, Medhat Shehata, Lucia Olexikova, Alexander V. Makarevich, Barbora Kulíková, Jaromír Vašíček, Rainer Hubmann, Peter Chrenek, Jozef Čurlej, Michal Kovac |
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Rok vydání: | 2018 |
Předmět: |
0106 biological sciences
Bioengineering Vimentin Bone Marrow Cells 01 natural sciences Cryopreservation 03 medical and health sciences SOX2 Antigens CD 010608 biotechnology medicine Animals CD90 RNA Messenger Molecular Biology 030304 developmental biology Biological Specimen Banks 0303 health sciences biology Reverse Transcriptase Polymerase Chain Reaction CD44 Mesenchymal stem cell Mesenchymal Stem Cells General Medicine Cell biology medicine.anatomical_structure Phenotype biology.protein CD146 Bone marrow Rabbits Biotechnology |
Zdroj: | New biotechnology. 54 |
ISSN: | 1876-4347 |
Popis: | 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. |
Databáze: | OpenAIRE |
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