Establishment of a tetracycline (Tet)-controlled <i>Amelx</i> expression system in MSCs.

Abstract

<p>(<b>a</b>): Schematic diagram depicting the procedure to establish a Tet-controlled <i>Amelx</i> expression system in MSCs. TetR: Tet repressor, TO: Tet operator, Dox: doxycycline (tetracycline derivative). (<b>b</b>): MSC colony (MSC-<i>TetR</i>) in culture medium containing 500 μg/mL geneticin 10 days after transduction with the pLenti3.3/<i>TetR</i> expression vector (bar; 60 μm). (<b>c</b>): Expression of TetR repressor gene in MSCs (without transduction) and MSCs-<i>TetR</i> was determined by RT-PCR. HEK293 cells subjected to the same transduction procedure (HEK-<i>TetR</i>) were used as a positive control. (<b>d, e</b>): MSCs-<i>TetR</i> were lentivirally transduced with the expression vector pLenti6.3/TO/V5/<i>Amelx</i> or plenti6.3/V5-GW/<i>GFP</i>. MSCs-<i>TetR</i>/<i>Amelx</i> (<b>d</b>) and MSCs-<i>TetR</i>/<i>GFP</i> (<b>e</b>) were selected by 10 μg/mL blastcidin S (bar; 200 μm).</p