Molecular Profiling and Gene Banking of Rabbit EPCs Derived from Two Biological Sources
Endothelial progenitor cells (EPCs) have been broadly studied for several years due to their outstanding regenerative potential. Moreover, these cells might be a valuable source of genetic information for the preservation of endangered animal species. However, a controversy regarding their character...
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2021-03-01
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| author | Jaromír Vašíček Andrej Baláži Miroslav Bauer Andrea Svoradová Mária Tirpáková Marián Tomka Peter Chrenek |
| author_facet | Jaromír Vašíček Andrej Baláži Miroslav Bauer Andrea Svoradová Mária Tirpáková Marián Tomka Peter Chrenek |
| author_sort | Jaromír Vašíček |
| collection | DOAJ |
| description | Endothelial progenitor cells (EPCs) have been broadly studied for several years due to their outstanding regenerative potential. Moreover, these cells might be a valuable source of genetic information for the preservation of endangered animal species. However, a controversy regarding their characterization still exists. The aim of this study was to isolate and compare the rabbit peripheral blood- and bone marrow-derived EPCs with human umbilical vein endothelial cells (HUVECs) in terms of their phenotype and morphology that could be affected by the passage number or cryopreservation as well as to assess their possible neuro-differentiation potential. Briefly, cells were isolated and cultured under standard endothelial conditions until passage 3. The morphological changes during the culture were monitored and each passage was analyzed for the typical phenotype using flow cytometry, quantitative real–time polymerase chain reaction (qPCR) and novel digital droplet PCR (ddPCR), and compared to HUVECs. The neurogenic differentiation was induced using a commercial kit. Rabbit cells were also cryopreserved for at least 3 months and then analyzed after thawing. According to the obtained results, both rabbit EPCs exhibit a spindle-shaped morphology and high proliferation rate. The both cell lines possess same stable phenotype: CD14<sup>−</sup>CD29<sup>+</sup>CD31<sup>−</sup>CD34<sup>−</sup>CD44<sup>+</sup>CD45<sup>−</sup>CD49f<sup>+</sup>CD73<sup>+</sup>CD90<sup>+</sup>CD105<sup>+</sup>CD133<sup>−</sup>CD146<sup>−</sup>CD166<sup>+</sup>VE-cadherin<sup>+</sup>VEGFR-2<sup>+</sup>SSEA-4<sup>+</sup>MSCA-1<sup>−</sup>vWF<sup>+</sup>eNOS<sup>+</sup>AcLDL<sup>+</sup>ALDH<sup>+</sup>vimentin<sup>+</sup>desmin<sup>+</sup>α-SMA<sup>+</sup>, slightly different from HUVECs. Moreover, both induced rabbit EPCs exhibit neuron-like morphological changes and expression of neuronal markers ENO2 and MAP2. In addition, cryopreserved rabbit cells maintained high viability (>85%) and endothelial phenotype after thawing. In conclusion, our findings suggest that cells expanded from the rabbit peripheral blood and bone marrow are of the endothelial origin with a stable marker expression and interesting proliferation and differentiation capacity. |
| first_indexed | 2024-03-09T05:34:10Z |
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| language | English |
| last_indexed | 2024-03-09T05:34:10Z |
| publishDate | 2021-03-01 |
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| spelling | doaj.art-15b4121f7de149919fe012515928d15f2023-12-03T12:30:20ZengMDPI AGGenes2073-44252021-03-0112336610.3390/genes12030366Molecular Profiling and Gene Banking of Rabbit EPCs Derived from Two Biological SourcesJaromír Vašíček0Andrej Baláži1Miroslav Bauer2Andrea Svoradová3Mária Tirpáková4Marián Tomka5Peter Chrenek6NPPC, Research Institute for Animal Production Nitra, Institute of Farm Animal Genetics and Reproduction, Hlohovecká 2, 951 41 Lužianky, SlovakiaNPPC, Research Institute for Animal Production Nitra, Institute of Farm Animal Genetics and Reproduction, Hlohovecká 2, 951 41 Lužianky, SlovakiaNPPC, Research Institute for Animal Production Nitra, Institute of Farm Animal Genetics and Reproduction, Hlohovecká 2, 951 41 Lužianky, SlovakiaNPPC, Research Institute for Animal Production Nitra, Institute of Farm Animal Genetics and Reproduction, Hlohovecká 2, 951 41 Lužianky, SlovakiaDepartment of Biochemistry and Biotechnology, Faculty of Biotechnology and Food Science, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2, 949 76 Nitra, SlovakiaNPPC, Research Institute for Animal Production Nitra, Institute of Farm Animal Genetics and Reproduction, Hlohovecká 2, 951 41 Lužianky, SlovakiaNPPC, Research Institute for Animal Production Nitra, Institute of Farm Animal Genetics and Reproduction, Hlohovecká 2, 951 41 Lužianky, SlovakiaEndothelial progenitor cells (EPCs) have been broadly studied for several years due to their outstanding regenerative potential. Moreover, these cells might be a valuable source of genetic information for the preservation of endangered animal species. However, a controversy regarding their characterization still exists. The aim of this study was to isolate and compare the rabbit peripheral blood- and bone marrow-derived EPCs with human umbilical vein endothelial cells (HUVECs) in terms of their phenotype and morphology that could be affected by the passage number or cryopreservation as well as to assess their possible neuro-differentiation potential. Briefly, cells were isolated and cultured under standard endothelial conditions until passage 3. The morphological changes during the culture were monitored and each passage was analyzed for the typical phenotype using flow cytometry, quantitative real–time polymerase chain reaction (qPCR) and novel digital droplet PCR (ddPCR), and compared to HUVECs. The neurogenic differentiation was induced using a commercial kit. Rabbit cells were also cryopreserved for at least 3 months and then analyzed after thawing. According to the obtained results, both rabbit EPCs exhibit a spindle-shaped morphology and high proliferation rate. The both cell lines possess same stable phenotype: CD14<sup>−</sup>CD29<sup>+</sup>CD31<sup>−</sup>CD34<sup>−</sup>CD44<sup>+</sup>CD45<sup>−</sup>CD49f<sup>+</sup>CD73<sup>+</sup>CD90<sup>+</sup>CD105<sup>+</sup>CD133<sup>−</sup>CD146<sup>−</sup>CD166<sup>+</sup>VE-cadherin<sup>+</sup>VEGFR-2<sup>+</sup>SSEA-4<sup>+</sup>MSCA-1<sup>−</sup>vWF<sup>+</sup>eNOS<sup>+</sup>AcLDL<sup>+</sup>ALDH<sup>+</sup>vimentin<sup>+</sup>desmin<sup>+</sup>α-SMA<sup>+</sup>, slightly different from HUVECs. Moreover, both induced rabbit EPCs exhibit neuron-like morphological changes and expression of neuronal markers ENO2 and MAP2. In addition, cryopreserved rabbit cells maintained high viability (>85%) and endothelial phenotype after thawing. In conclusion, our findings suggest that cells expanded from the rabbit peripheral blood and bone marrow are of the endothelial origin with a stable marker expression and interesting proliferation and differentiation capacity.https://www.mdpi.com/2073-4425/12/3/366rabbitperipheral bloodbone marrowendothelial progenitor cellsHUVECsflow cytometry |
| spellingShingle | Jaromír Vašíček Andrej Baláži Miroslav Bauer Andrea Svoradová Mária Tirpáková Marián Tomka Peter Chrenek Molecular Profiling and Gene Banking of Rabbit EPCs Derived from Two Biological Sources Genes rabbit peripheral blood bone marrow endothelial progenitor cells HUVECs flow cytometry |
| title | Molecular Profiling and Gene Banking of Rabbit EPCs Derived from Two Biological Sources |
| title_full | Molecular Profiling and Gene Banking of Rabbit EPCs Derived from Two Biological Sources |
| title_fullStr | Molecular Profiling and Gene Banking of Rabbit EPCs Derived from Two Biological Sources |
| title_full_unstemmed | Molecular Profiling and Gene Banking of Rabbit EPCs Derived from Two Biological Sources |
| title_short | Molecular Profiling and Gene Banking of Rabbit EPCs Derived from Two Biological Sources |
| title_sort | molecular profiling and gene banking of rabbit epcs derived from two biological sources |
| topic | rabbit peripheral blood bone marrow endothelial progenitor cells HUVECs flow cytometry |
| url | https://www.mdpi.com/2073-4425/12/3/366 |
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