Dmem f12

The Bombyx mori silkworm fibers were obtained from China (Jiuyuan mulberry silk, Hangzhou, China). For silk degumming, Na₂CO₃ (Sigma-Aldrich, St. Louis, MO, USA) was used. LiBr (Sigma-Aldrich, St. Louis, MO, USA) was used to dissolve the degummed silk fibers, and cellulose membranes (3500 MWCO, Thermo Scientific, Rockford, IL, USA) were used to dialyze the solution. Gelatin (Sinopharm, Peking, China) solution was used to mix with CSF and form hydrogel, chondrocytes (YaJi Biological, Shanghai, China) were cultured with the mixture of DMEM-F12 (Meilunbio, Dalian, China), fetal bovine serum (Sangon Biotech, Shanghai, China), and antibiotic/antimycotic (Sangon Biotech, Shanghai, China). The hydrogels with encapsulated cells were stained with a live/dead Viability Kit (Life Technologies, Grand Island, NY, USA).

The human GC tumor-derived cell line KGN was purchased from CELLCOOK biotechnology. Cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM)/F-12 (Meilunbio, China) supplemented with 10% fetal bovine serum (Gibco, Life Technologies, Grand Island, NY) and 100 units/mL penicillin and streptomycin (Invitrogen, Waltham, MA, United States) at 37°C with 5 % CO₂. Lentivirus was purchased from Shanghai GeneChem Co., LTD. Cells were treated with si-miR93-5p (silencing of miR-93-5p), si-NC, oe-miR93-5p (overexpression of miR-93-5p) Lentivirus and oe-NC. A fluorescence microscope with a digital camera (Olympus, Tokyo, Japan) was used to observe the fluorescence intensity of the cells. After 48 h of infection, the cells were selected with 5mg/mL puromycin (Invitrogen, United States). For inhibitor experiment, NF-κB inhibitor (BAY 11-7082) was purchased from MedChemExpress (MCE, China) and KGN cells were treated with 5 μM BAY 11-7082 for 24 h. Ferroptosis inhibitor (Fer-1) and ferroptosis inducer (erastin) were purchased from Good Laboratory Practice Bioscience (GLPBio, China).

The follicular fluid was pooled and centrifuged at 3000 rpm for 10 min. Then the pellets were resuspended in phosphate-buffered saline (Servicebio, Wuhan), and the suspension was slowly and carefully transferred to the same amount of FICOLL separation liquid surface and centrifuged for 5 min at 2500 rpm. After the delamination was obvious, the middle GCs layer was carefully sucked out and washed once with Dulbecco’s modified Eagle’s medium/nutrient mixture F-12 (DMEM-F12). All the GCs were cultured in DMEM-F12 (meilunbio, Dalian) containing 10% fetal bovine serum (Every Green, Zhejiang) and 1% penicillin–streptomycin-neomycin (Servicebio, Wuhan). The cells were placed in an incubator containing 5% CO₂ at 37 °C.

The brown preadipocytes were isolated from goat perirenal BAT at 1 day after birth. After washing with phosphate-buffered saline (PBS), the BAT was digested with type Ⅱ-collagen enzyme (2 mg/mL) (BioFrox, Guangzhou, China) for 25 min at 37 °C, followed by filtration through a 70 µm filter. After centrifugation at 1500× g for 5 min, we added the medium containing 10% FBS (Gibco, Emeryville, CA, USA), streptomycin, penicillin, and DMEM/F12 (Meilunbio, Dalian, China), and transferred it to the culture bottle under conditions of 5% CO₂ and 37 °C.
Primary white adipocytes were obtained from the perirenal WAT at 30 days after birth. Brown and white adipocytes were cultured in media supplemented with 10% FBS, 860 nM insulin (Solarbio, Beijing, China), 1 nM triiodothyronine(T3) (Selleck, Houston, TX, USA), 1 μM rosiglitazone (Selleck, TX, USA), 1 µM dexamethasone (MCE, Shanghai, China), and 0.5 mM IBMX (Sigma, Marlborough, MA, USA) for 2 days. Cells were then treated in maintenance media (2% FBS, 860 nM insulin, and 1 nM T3) for 6 days.

Human trophoblast cell lines (Jeg3, HTR8) and human decidual stromal cell line (hESC) were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Jeg3 and HTR8 cells were cultured in a 5% humidified carbon dioxide atmosphere at 37 °C in Dulbecco’s modified Eagle’s medium (DMEM)/F-12 (Gibco, Life Technologies, Grand Island, NY, USA) with 10% foetal bovine serum (Gibco, Life Technologies, Grand Island, NY, USA), 50 mg/mL streptomycin, and 50 U/mL penicillin. Jeg3 is the only trophoblast cell line that expresses human leukocyte antigen G (HLA-G), which is gradually expressed during development from CTBs to EVTs.
hESCs were cultured in a 5% humidified carbon dioxide atmosphere at 37 °C in phenol red-free Dulbecco’s modified Eagle’s medium (DMEM)/F-12 (Meilunbio, Dalian, China) with 10% foetal bovine serum, 50 mg/mL streptomycin, and 50 U/mL penicillin. One micromolar medroxyprogesterone-17-acetate (MPA) (HY-B0469, MedChemExpress, NJ, USA) and 0.5 mM N6,20-O-dibutyryladenosine cAMP sodium salt (db-cAMP) (HY-B0764, MedChemExpress, Monmouth Junction, NJ, USA) were added to the culture for 6 days to induce hESC decidualization in vitro [14 (link)].

Mouse hepatocyte AML12 cells were bought from the ATCC. They were cultured in Dulbecco’s Modified Eagle Medium/nutrient mixture F-12 (DMEM-F12; Meilun Bio, Dalian, China) with 10% fetal bovine serum, 100 U/ml penicillin/0.1 mg/ml streptomycin (Biosharp, Hefei, China), 1% insulin-transferrin-selenium (Thermo Fisher Scientific, Waltham, MA, USA), and 40 ng/ml dexamethasone (Beyotime). Human hepatocyte MIHA cells, kindly provided by Dr. J. R. Chowdhury (Albert Einstein College of Medicine, New York, NY, USA), were cultured in DMEM (Gibco, Shanghai, China) with 10% fetal bovine serum and 100 U/ml penicillin/0.1 mg/ml streptomycin. All cells were maintained in an incubator at 5% CO₂ and 37 °C.