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. 2019 Apr 17;10:248. doi: 10.3389/fendo.2019.00248

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Copyright © 2019 Bjelobaba, Janjic, Prévide, Abebe, Kucka and Stojilkovic.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Culturing of dispersed pituitary cells stimulates Spp1 expression. (A) The time course of basal Spp1 expression in cultured pituitary cells. Cells were derived from 7-week-old female and male rats. Zero indicates Spp1 expression levels immediately after cell dispersion. Cells were cultured in medium 199 containing horse serum. (B) Opposite effects of dispersion and culturing of pituitary cells from females on Spp1 and Gnrhr expression. In (A) and (B), cells were continuously cultured in medium 199 containing horse serum without replacement of old medium with fresh. (C,D) Effects of replacement of culturing medium on rate of basal Spp1 (C) and Gnrhr (D) expression. Old media were replaced with fresh media after 12, 36, and 60 h incubation and cells were collected for mRNA extraction immediately after dispersion, and 24, 48, and 72 h after dispersion.