Mycoplasma detection kit

The ccRCC cell lines and renal cell lines used in the present study were obtained from the Cell Bank of the Type Culture Collection of the Chinese Academy of Sciences in 2018. HK-2 cells were maintained in high-glucose Dulbecco’s Modified Eagle’s Medium (DMEM) (Gibco, 11995–065). ACHN cells were cultured in Minimum Essential Medium (Gibco, 11095–080), and 786-O, 769-P and OS-RC-2 cells were maintained in RPMI-1640 medium (Gibco, 22400–089). The culture media of all cell lines were supplemented with fetal bovine serum (FBS, 10%, Gibco) and 1% penicillin/streptomycin (Gibco). ccRCC cell lines were cultured at 37°C in 5% Co₂. Sunitinib- and pazopanib-resistant 786-O cell lines were maintained in RPMI-1640 medium supplemented with 10% (v/v) FBS and 10 µM sunitinib or 8 µM pazopanib. All cell lines in the present study were authenticated by short tandem repeat (STR) profiling and examined for mycoplasma contamination using a Mycoplasma Detection Kit (Selleck Chemicals), and the most recent tests were conducted in June 2019. All cell lines used in the study were cultured within 40 passages.

RCC cell lines used in this study were obtained from the Cell Bank of the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China) in 2017. HK-2 cells were maintained in DMEM, High Glucose medium (Gibco, Waltham, MA, USA). A498 and ACHN cells were cultured in Minimum Essential Medium (Gibco). 786-O cells were maintained in RPMI-1640 medium (Gibco). Caki-1cells were maintained in McCoy's 5A Medium (Gibco). The culture media of all cell lines were supplemented with fetal bovine serum (FBS, 10%, Gibco), 1% penicillin/streptomycin (Gibco). RCC cell lines were cultured at 37 °C in 5% Co₂. Sunitinib-resistant 786-O and 769-p cell lines (786-O-SR and 769-p-SR) were continuously exposed to increasing dose of Sunitinib (Selleck, USA) for about 12 weeks. The starting dose was 5 μM and this was increased to 10 μM after 4 weeks, to 15 μM after another 4 weeks and continued at 15 μM for the last 4 weeks. The established resistant 786-O and 769-P cell lines were then maintained in DMEM medium with 10% (v/v) FBS and 10 μM Sunitinib.
The cell lines in this study were authenticated by short tandem repeat (STR) profiling and detected for mycoplasma contamination using a Mycoplasma Detection Kit (Selleck Chemicals), and the most recent tests were conducted in September 2018. All cell lines used in this study were cultured within 40 passages.