An efficient procedure of isolation, cultivation and identification of bone marrow mesenchymal stem cells
Background: Bone marrow mesenchymal stem cells (MSC) have a wide application in domain of Regenerative Medicine. Of a great importance is utilization of a suitable bone marrow extraction technique that can provide a sufficient number of MSC to perform laboratory tests without seriously affecting the...
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| Format: | Article |
| Language: | English |
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Scientific Medical Association of Moldova
2019-03-01
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| Series: | The Moldovan Medical Journal |
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| Online Access: | http://moldmedjournal.md/wp-content/uploads/2019/03/moldovan-med-j-2019-62-1-cobzac-et-al-full-text.pdf |
| _version_ | 1818083246361018368 |
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| author | Vitalie Cobzac Andrei Mostovei Mariana Jian Viorel Nacu |
| author_facet | Vitalie Cobzac Andrei Mostovei Mariana Jian Viorel Nacu |
| author_sort | Vitalie Cobzac |
| collection | DOAJ |
| description | Background: Bone marrow mesenchymal stem cells (MSC) have a wide application in domain of Regenerative Medicine. Of a great importance is utilization of a suitable bone marrow extraction technique that can provide a sufficient number of MSC to perform laboratory tests without seriously affecting the health of the laboratory animal. At the same time, before using in researches and clinical application, the MSC needs to be identified. Material and methods: The study was conducted in rabbits (n = 9), in which, from one iliac bone, by aspiration were taken 3.39 ± 1.27 ml of bone marrow. The nucleated bone marrow cells were separated through centrifugation using concentration gradient. The specific for stem cells culture medium was used, and MSC were multiplied during 2 passages. From the obtained MSC, 1x106 cells were subject to differentiation by chondrocytes lineage for other 20 days. The obtained chondrocytes aggregates were morphologically examined by Hematoxylin–Eosin staining and specific cartilage staining with Safranin O and Toluidine blue/fast green. Results: There was a strong correlation between the volume of collected bone marrow and the time required to achieve a 70-80% of MSC confluence (p=0.01). Also, the MSC isolated from bone marrow extracted from rabbit iliac bone were differentiated successful on chondrocyte line in all cases, confirmed through the specific cartilage staining with Safranin O and Toluidine blue/fast green (p<0,001). Conclusions: The volume of 3.39 ± 1.27 ml of bone marrow, harvested from rabbit iliac bone is sufficient to obtain a large number of MSC for the laboratory tests in vitro and in vivo. As a standard method for MSC identification could be used just the capability of the cells to differentiate in the specialized cell, including chondrocytes. |
| first_indexed | 2024-12-10T19:34:57Z |
| format | Article |
| id | doaj.art-2a2a0b720f954a3d89dbf8c31121ac32 |
| institution | Directory Open Access Journal |
| issn | 2537-6373 2537-6381 |
| language | English |
| last_indexed | 2024-12-10T19:34:57Z |
| publishDate | 2019-03-01 |
| publisher | Scientific Medical Association of Moldova |
| record_format | Article |
| series | The Moldovan Medical Journal |
| spelling | doaj.art-2a2a0b720f954a3d89dbf8c31121ac322022-12-22T01:36:10ZengScientific Medical Association of MoldovaThe Moldovan Medical Journal2537-63732537-63812019-03-01621354010.5281/zenodo.2590011An efficient procedure of isolation, cultivation and identification of bone marrow mesenchymal stem cellsVitalie Cobzac0Andrei Mostovei1Mariana Jian2Viorel Nacu3Tissue Engineering and Cells Cultures Laboratory, Nicolae Testemitsanu State University of Medicine and Pharmacy, Chisinau, the Republic of MoldovaTissue Engineering and Cells Cultures Laboratory, Nicolae Testemitsanu State University of Medicine and Pharmacy, Chisinau, the Republic of MoldovaTissue Engineering and Cells Cultures Laboratory, Nicolae Testemitsanu State University of Medicine and Pharmacy, Chisinau, the Republic of MoldovaTissue Engineering and Cells Cultures Laboratory, Nicolae Testemitsanu State University of Medicine and Pharmacy, Chisinau, the Republic of MoldovaBackground: Bone marrow mesenchymal stem cells (MSC) have a wide application in domain of Regenerative Medicine. Of a great importance is utilization of a suitable bone marrow extraction technique that can provide a sufficient number of MSC to perform laboratory tests without seriously affecting the health of the laboratory animal. At the same time, before using in researches and clinical application, the MSC needs to be identified. Material and methods: The study was conducted in rabbits (n = 9), in which, from one iliac bone, by aspiration were taken 3.39 ± 1.27 ml of bone marrow. The nucleated bone marrow cells were separated through centrifugation using concentration gradient. The specific for stem cells culture medium was used, and MSC were multiplied during 2 passages. From the obtained MSC, 1x106 cells were subject to differentiation by chondrocytes lineage for other 20 days. The obtained chondrocytes aggregates were morphologically examined by Hematoxylin–Eosin staining and specific cartilage staining with Safranin O and Toluidine blue/fast green. Results: There was a strong correlation between the volume of collected bone marrow and the time required to achieve a 70-80% of MSC confluence (p=0.01). Also, the MSC isolated from bone marrow extracted from rabbit iliac bone were differentiated successful on chondrocyte line in all cases, confirmed through the specific cartilage staining with Safranin O and Toluidine blue/fast green (p<0,001). Conclusions: The volume of 3.39 ± 1.27 ml of bone marrow, harvested from rabbit iliac bone is sufficient to obtain a large number of MSC for the laboratory tests in vitro and in vivo. As a standard method for MSC identification could be used just the capability of the cells to differentiate in the specialized cell, including chondrocytes.http://moldmedjournal.md/wp-content/uploads/2019/03/moldovan-med-j-2019-62-1-cobzac-et-al-full-text.pdfmesenchymal stem cellsbone marrowrabbitscellular identificationiliac boneautocells |
| spellingShingle | Vitalie Cobzac Andrei Mostovei Mariana Jian Viorel Nacu An efficient procedure of isolation, cultivation and identification of bone marrow mesenchymal stem cells The Moldovan Medical Journal mesenchymal stem cells bone marrow rabbits cellular identification iliac bone autocells |
| title | An efficient procedure of isolation, cultivation and identification of bone marrow mesenchymal stem cells |
| title_full | An efficient procedure of isolation, cultivation and identification of bone marrow mesenchymal stem cells |
| title_fullStr | An efficient procedure of isolation, cultivation and identification of bone marrow mesenchymal stem cells |
| title_full_unstemmed | An efficient procedure of isolation, cultivation and identification of bone marrow mesenchymal stem cells |
| title_short | An efficient procedure of isolation, cultivation and identification of bone marrow mesenchymal stem cells |
| title_sort | efficient procedure of isolation cultivation and identification of bone marrow mesenchymal stem cells |
| topic | mesenchymal stem cells bone marrow rabbits cellular identification iliac bone autocells |
| url | http://moldmedjournal.md/wp-content/uploads/2019/03/moldovan-med-j-2019-62-1-cobzac-et-al-full-text.pdf |
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