Combined approach for characterization and quality assessment of rabbit bone marrow-derived mesenchymal stem cells intended for gene banking

被引：14

作者：

Vasicek, Jaromir ^{[1
,2
,3
]}

Kovac, Michal ^{[3
]}

Balazi, Andrej ^{[1
]}

Kulikova, Barbora ^{[1
]}

Tomkova, Maria ^{[3
]}

Olexikova, Lucia ^{[1
]}

Curlej, Jozef ^{[3
]}

Bauer, Miroslav ^{[1
,4
]}

Schnabl, Susanne ^{[5
]}

Hilgarth, Martin ^{[5
]}

Hubmann, Rainer ^{[5
]}

Shehata, Medhat ^{[5
]}

Makarevich, Alexander V. ^{[1
]}

Chrenek, Peter ^{[1
,3
,6
]}

机构：

[1] NPPC Res Inst Anim Prod Nitra, Hlohovecka 2, Luzianky 95141, Slovakia

[2] Slovak Univ Agr, Res Ctr AgroBioTech, Tr A Hlinku 2, Nitra 94976, Slovakia

[3] Slovak Univ Agr, Fac Biotechnol & Food Sci, Tr A Hlinku 2, Nitra 94976, Slovakia

[4] Constantine Philosopher Univ Nitra, Fac Nat Sci, Nabrezie Mladeze 91, Nitra 94974, Slovakia

[5] Med Univ Vienna, Div Haematol & Haemostaseol, Internal Med 1, Drug & Target Screening Unit DTSU, Waehringer Guertel 18-20, A-1090 Vienna, Austria

[6] UTP Univ Sci & Technol, Fac Anim Breeding & Biol, Mazowiecka 28, PL-85084 Bydgoszcz, Poland

来源：

NEW BIOTECHNOLOGY | 2020年 / 54卷

关键词：

Rabbit; MSCs; Flow cytometry; PCR; TEM; Cryopreservation; OSTEOGENIC DIFFERENTIATION; PROGENITOR CELLS; FETAL ADNEXA; EXPRESSION; FLUID; BLOOD; CRYOPRESERVATION; IDENTIFICATION; ESTABLISHMENT; TISSUE;

D O I：

10.1016/j.nbt.2019.08.001

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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 alpha-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.

引用

页码：1 / 12

页数：12

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