CHARACTERIZATION OF CULTURED SERTOLI CELLS UNDER HIGH-TEMPERATURE AND HYPOXIC CONDITIONS
A.Yu. Kulibin,1 E.A. Malolina 2
1 Koltzov Institute of Developmental Biology, RAS and 2 D.I. Ivanovsky Institute of Virology Ministry of Health
and Social Development of the Russian Federation, Moscow;
1 e-mail: Kulibin.A.BKRJ@gmail.com
Sertoli cells (SCs) isolated from adult C57Bl/6 mice were characterized under four different cell culture conditions: standard conditions (34 °Ñ, 21 % O2 - 34_21),
high-temperature conditions (37 °Ñ, 21 % O2 - 37_21), hypoxic conditions (34 °Ñ, 5 % O2 - 34_5), and combination of these conditions
(37 °Ñ, 5 % O2 - 37_5). Proliferation and viability were promoted when SCs were grown under hypoxia: 28.5 and 24.6 % of SCs were BrdU-positive at the peak
of proliferation, 92.7 and 92.7 % of SCs were viable after 15 days in culture at 34_5 and 37_5, respectively, versus 20.2 and 88.9 % at 34_21, respectively. In SCs grown under
high-temperature conditions proliferation was slightly increased, but viability was decreased: 23.1 % of SCs were BrdU-positive, and only 74.9 % of SCs were viable at 37_21.
At the same time cultivation of SCs at 37 °Ñ promoted their dedifferentiation: after 15 days in culture 98.8 and 98.6 % of cells at 37_5 and 37_21, respectively, expressed a marker
of immature SCs - cytokeratin-18, compared to 26.5 % at 34_5 and 6.6 % at 34_21. Expression of Wt1, a transcription factor controlling cell-cell junction formation and germ
cell development, disappeared in most cells after 3 days in culture under all culture conditions. However, SCs forming colonies restored Wt1 expression at day 15 in culture under
high-temperature conditions: 59.1 and 29.5 % of SCs were Wt1-positive at 37_21 and 37_5, respectively, versus 11.1 and 3.6 % at 34_21 and 34_5, respectively. Cultured SCs
expressed other SC markers (vimentin, clusterin, Gata-4) under all culture conditions. Our results show that cultured SCs may be useful for reproductive biology and regenerative
medicine.
Key words: Sertoli cells, hypoxia, high-temperature conditions, cytokeratin-18, Wt1
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