Mycoalert lt07 318

D341 (ATCC^® HTB-187TM, RRID:CVCL_0018) and HEK293T (ATCC^® CRL-3216, RRID:CVCL_0063) were purchased from ATCC, and D425 (SCC290, RRID:CVCL_1275) was purchased from Sigma-Millipore (Sigma-Aldrich, St. Louis, MO, USA) [54 (link),55 (link),56 (link)]. Human medulloblastoma cells were grown in Minimum Essential Media (MEM; Gibco 11095; Thermo Fisher Scientific, Waltham, MA, USA) plus 15% fetal bovine serum (FBS; Gibco 26140079; Thermo Fisher Scientific, Waltham, MA, USA), 100 mg/mL penicillin G, and 100 lg/mL streptomycin (Gibco 15140122). HEK293T cells were grown in Dulbecco’s Modified Eagle’s Medium (DMEM; Gibco 11965092) supplemented with 10% FBS (Gibco 26140079), 100 mg/mL penicillin G, and 100 lg/mL streptomycin (Gibco 15140122; Thermo Fisher Scientific, Waltham, MA, USA). All cell lines were cultured at 37 °C in a humidified environment with 5% CO₂ and confirmed mycoplasma negative (Lonza MycoAlert LT07-318; Basel, Switzerland) (most recently confirmed negative in January 2024).

The KGN cell line [88 (link)] was a kind gift from André Pèlegrin (IRCM, Institut de Recherche en Cancérologie de Montpellier, Montpellier, France). KGN cells showed a pattern similar to that of steroidogenesis in human granulosa cells, thus allowing for the analysis of naturally occurring steroidogenesis in human granulosa cells [88 (link)]. The Fas-mediated apoptosis of KGN was also observed, which mimicked the physiological regulation of apoptosis in normal human granulosa cells [84 (link)]. Based on these findings, this cell line is considered to be a very useful model for understanding the regulation of steroidogenesis, cell growth, and apoptosis in human granulosa cells [88 (link)]. Cells were grown in DMEM/F-12 medium containing 10% heat-inactivated fetal bovine serum and 100 IU/mL penicillin/streptomycin (i.e., culture medium). Cells were grown at 37 °C in a humidified atmosphere with 5% CO₂, and medium was replaced twice per week. Cells were harvested twice per week with 0.5 mg/ml trypsin/0.2 mg/mL EDTA. KGN had a relatively long population doubling time of about 46.4 h [84 (link)]. All culture media and supplements were purchased from Life Technologies Inc. (Life Technologies Inc., Paisley, UK). The absence of mycoplasma was tested every four weeks (MycoAlert LT07-318, Lonza, Walkersville, MD, USA).

The human COV434^{41 (link)} and KGN^{42 (link)} cell lines were kind gifts from Dr. PI Schrier (Department of Clinical Oncology, Leiden University Medical Center, Netherland) and Dr T Yanase (Kyushu University, Fukuoka, Japan), respectively. The human high grade serous ovarian cancer cell lines SKOV3 and NIH-OVCAR8 were from ATCC (ATCC HTB-77) and from the Division of Cancer Treatment and Diagnosis, NCI, Frederick, MD, USA, respectively. Cells were grown in DMEM/F-12 medium without red phenol containing 10% heat-inactivated FBS. COV434-AMHRII and SKOV3-AMHRII cells were supplemented with 0.33 mg/ml geneticin (InvivoGen, ant-gn-1). Cells were grown at 37 °C in a humidified atmosphere with 5% CO₂, and medium was replaced twice per week. Cells were harvested with 0.5 mg/ml trypsin/0.2 mg/ml EDTA. All culture media and supplements were purchased from Life Technologies. Inc. (Gibco BRL). HEK293T cells, used for antibody production by the GenAc platform at IRCM, were grown in DMEM/F-12 with phenol red and 10% heat-inactivated FBS. The absence of Mycoplasma was tested every other week (MycoAlert LT07-318 & LT07-518, Lonza Walkersville, USA). The COV434-AMHRII and SKOV3-AMHRII cell lines were generated by transfection of the cDNA encoding full-length human AMHRII according to Kersual et al.^{43 (link)}.