Stable transfection cell lines

In order to induce upregulation of BMP2 and Smad1 levels in BMSCs, overexpressing lentivirus vectors targeting the rat BMP2 (OE-BMP2) and Smad1 (OE-Smad1) genes were designed and synthesized by GenePharma Co., Ltd. Briefly, the BMSCs (5×10⁵) were seeded in 6-well plates and cultured at 37°C overnight. Next, the cells were infected with OE-BMP2, OE-Smad1 or OE-NC (serving as the negative control vector) using 5 µg/ml polybrene (Hanbio Biotechnology Co., Ltd.), followed by incubation with 100 µg/ml G418 or 7 µg/ml puromycin for 14 days to select the stably infected cell lines, respectively.