Biopolym. Cell. 2022; 38(4):224-230.
Molecular and Cell Biotechnologies
Isolation and cultivation of MSC-like population from the rabbit amniotic membrane
- State institution “O. S. Kolomiychenko Institute of Otolaryngology Of National Academy of Medical Sciences of Ukraine”
3, Zoologichna, Kyiv, Ukraine, 02000 - State Institute of Genetic and Regenerative Medicine, NAMS of Ukraine
67, Vyshhorodska Str., Kyiv, Ukraine, 04114
Abstract
Despite the advantages of studying the therapeutic potential of autologous and allogeneic mesenchymal stem cells (MSCs) on the rabbit model, the cultivation of rabbit amnion MSCs has not been described in the literature. Aim. To study the possibility of isolation, multiplication, and cryopreservation of amnion-derived rabbit MSCs. Methods. The amniotic membrane was obtained surgically. Both enzymatic and explant methods of cell isolation have been tested. The obtained primary cultures were passed, and their proliferation capacity and morphology characteristics were studied. Results. The viable clones were obtained on the 2nd day of cultivation using an enzymatic method. At all passages, the cells showed adhesion to the culture plastic and fibroblast-like morphology. At the first and second passages, after 7 days of cultivation, the population doubling occurred 4.4 and 4.3 times, respectively. Evaluation of the viability after cryopreservation showed that after thawing, more than 90 % of cells were alive, and 4.3 doubling occurred in 7 days of cultivation. No cells with the atypical phenotype were detected during cultivation. Conclusions. The methods of extraction, multiplication, and cryopreservation of MSCs-like rabbit amnion cells were optimized. This can promote further studies of the MSCs characteristics and regenerative potential.
Keywords: rabbit amnion, MSC, primary culture
Full text: (PDF, in English)
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