BIOLOGICAL PECULIARITIES OF ADIPOSE TISSUE-DERIVED MESENCHYMAL STEM CELLS AT DIFFERENT PASSAGES OF CULTIVATION
The studies were conducted on 2–3-months-old males of C57BL/6 mice weighing 20–24 g. Obtaining and operating with adipose tissue-derived mesenchymal stem cell (MSC) culture was performed in a sterile laminar box under conditions of asepsis and antiseptics. The adipose tissue-derived MSC of the 2, 4,...
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| Format: | Article |
| Language: | English |
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National University of Life and Environmental Sciences of Ukraine
2021-11-01
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| Series: | Ukrainian Journal of Veterinary Sciences |
| Subjects: | |
| Online Access: | https://veterinaryscience.com.ua/en/journals/tom-12-4-2021/biologichni-osoblivosti-myezyenkhimalnikh-stovburovikh-klitin-iz-zhirovoyi-tkanini-za-riznikh-pasazhiv |
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| author | Larysa Kladnytska Anatolii Mazurkevych S. V. VELYCHKO L. V. GARMANCHUK Mykola Maliuk Tetiana Mazurkevych Vitalii Kovpak T. V. KOZYTSKA Yu. O. KHARKEVYCH Roman Bokotko T. L. SAVCHUK |
| author_facet | Larysa Kladnytska Anatolii Mazurkevych S. V. VELYCHKO L. V. GARMANCHUK Mykola Maliuk Tetiana Mazurkevych Vitalii Kovpak T. V. KOZYTSKA Yu. O. KHARKEVYCH Roman Bokotko T. L. SAVCHUK |
| author_sort | Larysa Kladnytska |
| collection | DOAJ |
| description | The studies were conducted on 2–3-months-old males of C57BL/6 mice weighing 20–24 g. Obtaining and operating with adipose tissue-derived mesenchymal stem cell (MSC) culture was performed in a sterile laminar box under conditions of asepsis and antiseptics. The adipose tissue-derived MSC of the 2, 4, 7 and 12 passages were analyzed. Morphometric analysis was performed using a light microscopy. Morphometric parameters such as cell and nucleus area or nuclear-cytoplasmic ratio were calculated using the Axiovision light microscope (Carl Zeiss, Germany) and Image J 1.45 software. Trypan blue dye used for investigation of the viability of MSC. The morphological characteristics of adipose tissue-derived MSC during the process of cultivation changes: at the first passages of cultivation, the cells are spindle-shaped with two, at least three, long cytoplasmic processes, which are located bipolar. Near the nucleus, the Golgi complex is clearly visible – a sign of active cells. At later passages, cells have a small cytoplasmic processes and the bipolar arrangement of processes changes by stellar arrangement. Golgi complex is also clearly visualized. The indicator of the nuclearcytoplasmic ratio in MSC from adipose tissue is significantly reduced at the 7th passage to 0.2189 ± 0.0122 (P < 0.01), and at the 12th passage to 0.1111 ± 0.0086 (P < 0.001) compared to the 2nd passage. The coefficient of proliferation of adipose tissue-derived MSC is significantly reduced at 12th passage. The viability of MSC from adipose tissue with an increasing of a number of passages significantly reduces and at the 12th passage of cultivation reaches 84.67 ± 1.36 (P < 0.05). The content of apoptotic cells that exhibited sensitivity to serum-free cultivation significantly increased at the 7th and 12th passages and was 21.33 ± 1.36 (P < 0.05) and 23.67 ± 0.97% (P < 0.05), respectively |
| first_indexed | 2024-04-25T01:37:08Z |
| format | Article |
| id | doaj.art-e55c473ab20b4ca4bf234168d3ebe75a |
| institution | Directory Open Access Journal |
| issn | 2663-967X 2663-9688 |
| language | English |
| last_indexed | 2024-04-25T01:37:08Z |
| publishDate | 2021-11-01 |
| publisher | National University of Life and Environmental Sciences of Ukraine |
| record_format | Article |
| series | Ukrainian Journal of Veterinary Sciences |
| spelling | doaj.art-e55c473ab20b4ca4bf234168d3ebe75a2024-03-08T09:18:37ZengNational University of Life and Environmental Sciences of UkraineUkrainian Journal of Veterinary Sciences2663-967X2663-96882021-11-01124798910.31548/ujvs2021.04.006BIOLOGICAL PECULIARITIES OF ADIPOSE TISSUE-DERIVED MESENCHYMAL STEM CELLS AT DIFFERENT PASSAGES OF CULTIVATIONLarysa Kladnytska0https://orcid.org/0000-0002-9360-0587Anatolii Mazurkevych1S. V. VELYCHKO2L. V. GARMANCHUK3Mykola Maliuk4https://orcid.org/0000-0003-3019-6035Tetiana Mazurkevych5https://orcid.org/0000-0002-1294-5939Vitalii Kovpak6https://orcid.org/0000-0003-2419-1246T. V. KOZYTSKA7Yu. O. KHARKEVYCH8Roman Bokotko9https://orcid.org/0000-0002-6217-5266T. L. SAVCHUK10National University of Life and Environmental Sciences of UkraineNational University of Life and Environmental Sciences of UkraineHospital of Veterinary MedicineTaras Shevchenko National University of Kyiv, Educational and Scientific Center Institute of Biology and MedicineNational University of Life and Environmental Sciences оf UkraineNational University of Life and Environmental Sciences of UkraineNational University of Life and Environmental Sciences of UkraineBogomolets National Medical UniversityNational University of Life and Environmental Sciences of UkraineNational University of Life and Environmental Sciences of UkraineNational University of Life and Environmental Sciences of UkraineThe studies were conducted on 2–3-months-old males of C57BL/6 mice weighing 20–24 g. Obtaining and operating with adipose tissue-derived mesenchymal stem cell (MSC) culture was performed in a sterile laminar box under conditions of asepsis and antiseptics. The adipose tissue-derived MSC of the 2, 4, 7 and 12 passages were analyzed. Morphometric analysis was performed using a light microscopy. Morphometric parameters such as cell and nucleus area or nuclear-cytoplasmic ratio were calculated using the Axiovision light microscope (Carl Zeiss, Germany) and Image J 1.45 software. Trypan blue dye used for investigation of the viability of MSC. The morphological characteristics of adipose tissue-derived MSC during the process of cultivation changes: at the first passages of cultivation, the cells are spindle-shaped with two, at least three, long cytoplasmic processes, which are located bipolar. Near the nucleus, the Golgi complex is clearly visible – a sign of active cells. At later passages, cells have a small cytoplasmic processes and the bipolar arrangement of processes changes by stellar arrangement. Golgi complex is also clearly visualized. The indicator of the nuclearcytoplasmic ratio in MSC from adipose tissue is significantly reduced at the 7th passage to 0.2189 ± 0.0122 (P < 0.01), and at the 12th passage to 0.1111 ± 0.0086 (P < 0.001) compared to the 2nd passage. The coefficient of proliferation of adipose tissue-derived MSC is significantly reduced at 12th passage. The viability of MSC from adipose tissue with an increasing of a number of passages significantly reduces and at the 12th passage of cultivation reaches 84.67 ± 1.36 (P < 0.05). The content of apoptotic cells that exhibited sensitivity to serum-free cultivation significantly increased at the 7th and 12th passages and was 21.33 ± 1.36 (P < 0.05) and 23.67 ± 0.97% (P < 0.05), respectivelyhttps://veterinaryscience.com.ua/en/journals/tom-12-4-2021/biologichni-osoblivosti-myezyenkhimalnikh-stovburovikh-klitin-iz-zhirovoyi-tkanini-za-riznikh-pasazhivadipose tissue mesenchymal stem cellsnucleusnuclear-cytoplasmic ratiocoefficient of proliferationviabilityapoptosisearly passageslate passages |
| spellingShingle | Larysa Kladnytska Anatolii Mazurkevych S. V. VELYCHKO L. V. GARMANCHUK Mykola Maliuk Tetiana Mazurkevych Vitalii Kovpak T. V. KOZYTSKA Yu. O. KHARKEVYCH Roman Bokotko T. L. SAVCHUK BIOLOGICAL PECULIARITIES OF ADIPOSE TISSUE-DERIVED MESENCHYMAL STEM CELLS AT DIFFERENT PASSAGES OF CULTIVATION Ukrainian Journal of Veterinary Sciences adipose tissue mesenchymal stem cells nucleus nuclear-cytoplasmic ratio coefficient of proliferation viability apoptosis early passages late passages |
| title | BIOLOGICAL PECULIARITIES OF ADIPOSE TISSUE-DERIVED MESENCHYMAL STEM CELLS AT DIFFERENT PASSAGES OF CULTIVATION |
| title_full | BIOLOGICAL PECULIARITIES OF ADIPOSE TISSUE-DERIVED MESENCHYMAL STEM CELLS AT DIFFERENT PASSAGES OF CULTIVATION |
| title_fullStr | BIOLOGICAL PECULIARITIES OF ADIPOSE TISSUE-DERIVED MESENCHYMAL STEM CELLS AT DIFFERENT PASSAGES OF CULTIVATION |
| title_full_unstemmed | BIOLOGICAL PECULIARITIES OF ADIPOSE TISSUE-DERIVED MESENCHYMAL STEM CELLS AT DIFFERENT PASSAGES OF CULTIVATION |
| title_short | BIOLOGICAL PECULIARITIES OF ADIPOSE TISSUE-DERIVED MESENCHYMAL STEM CELLS AT DIFFERENT PASSAGES OF CULTIVATION |
| title_sort | biological peculiarities of adipose tissue derived mesenchymal stem cells at different passages of cultivation |
| topic | adipose tissue mesenchymal stem cells nucleus nuclear-cytoplasmic ratio coefficient of proliferation viability apoptosis early passages late passages |
| url | https://veterinaryscience.com.ua/en/journals/tom-12-4-2021/biologichni-osoblivosti-myezyenkhimalnikh-stovburovikh-klitin-iz-zhirovoyi-tkanini-za-riznikh-pasazhiv |
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