Morphological peculiarites and functional activity of adipose-derived mesenchimal stem cells during in vitro cultivation conditions


  • L. V. Kladnytska National University of Life and Environmental Sciences of Ukraine, Kyiv, Ukraine http://orcid.org/0000-0002-9360-0587
  • A. I. Mazurkevych National University of Life and Environmental Sciences of Ukraine, Kyiv, Ukraine http://orcid.org/0000-0002-2409-7703
  • V. T. Khomych National University of Life and Environmental Sciences of Ukraine, Kyiv, Ukraine
  • T. A. Mazurkevych National University of Life and Environmental Sciences of Ukraine, Kyiv, Ukraine https://orcid.org/0000-0002-1294-5939
  • Z. G. Stegney National University of Life and Environmental Sciences of Ukraine, Kyiv, Ukraine https://orcid.org/0000-0002-0801-0751
  • M. O. Maluk National University of Life and Environmental Sciences of Ukraine, Kyiv, Ukraine http://orcid.org/0000-0003-3019-6035
  • L. V. Garmanchuk National Taras Shevchenko University, Educational and Scientific Center “Institute of Biology and Medicine”, Kyiv, Ukraine
  • S. V. Velychko Hospital of Veterinary Medicine, Kyiv, Ukraine http://orcid.org/0000-0003-1842-5114
  • V. B. Danilov National University of Life and Environmental Sciences of Ukraine, Kyiv, Ukraine http://orcid.org/0000-0002-2897-5235
  • Y. O. Kharkevych National University of Life and Environmental Sciences of Ukraine, Kyiv, Ukraine
  • D. V. Shelest National Taras Shevchenko University, Educational and Scientific Center “Institute of Biology and Medicine”, Kyiv, Ukraine http://orcid.org/0000-0002-6920-5640
  • V. S. Velychko National University of Life and Environmental Sciences of Ukraine, Kyiv, Ukraine http://orcid.org/0000-0002-9252-7365
  • I. A. Stupak National Taras Shevchenko University, Educational and Scientific Center “Institute of Biology and Medicine”, Kyiv, Ukraine
Keywords: Abrus precatorius, Vaccine, Immunoinhibition, Canine parvovirus

Abstract

The studies were conducted on 2-3-months-old males of C57BL/6 mice weighing 2024 g. Obtaining and cultivating of adipose-derived mesenchimal stem cells (AD MSCs) were carried out in a sterile laminar box with compliance of conditions of asepsis and antiseptics. AD MSCs 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 (NCR) were calculated using the Axiovision light microscope (Carl Zeiss, Germany) and ImageJ 1.45 software. Trypan blue dye used for investigation of the viability of MSC. The morphological characteristics of mesenchymal stem cells from adipose tissue during the process of cultivation changes: at the first passages of cultivation, the cells are spindle-shaped with two, at least three, long long cytoplasmic processes, 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 nuclear-cytoplasmic ratio in MSC from adipose tissue is significantly reduced at 7 passage to 0.2189 ± 0.0122 (P < 0.01), and at 12 passage to 0.1111 ± 0.0086 (P < 0.001) compared to the 2 passage. The coefficient of proliferation of MSC from adipose tissue is significantly reduced at 12th passage. The viability of mesenchymal stem cells 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 significantly increased at 7 and 12 passages and was respectively 21.33 ± 1.36 (P < 0.05) and 23.67 ± 0.97% (P < 0.05).

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Published
2018-12-10
How to Cite
Kladnytska, L., Mazurkevych, A., Khomych, V., Mazurkevych, T., Stegney, Z., Maluk, M., Garmanchuk, L., Velychko, S., Danilov, V., Kharkevych, Y., Shelest, D., Velychko, V., & Stupak, I. (2018). Morphological peculiarites and functional activity of adipose-derived mesenchimal stem cells during in vitro cultivation conditions. Scientific Messenger of LNU of Veterinary Medicine and Biotechnologies, 20(92), 79-82. https://doi.org/10.32718/nvlvet9216