Mammalian cell culture media BOSC23

Cells are sourced from various tissues to grow them in in-vitro conditions. Therefore, cell specific nutrients are important for their survival, maintenance and growth. Determining the appropriate cell culture media is a challenge if you are growing a cell line or a microorganism for the first time. Established cell lines, primary cells, stem cells, bacteria and Yeast all require varied nutrients from basic to complex. Based on the cell type, one can easy find what media and nutrients your peers have used before you try to reinvent the wheel.

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DMEM–Dulbecco's Modified Eagle Medium

Thermo Fisher Scientific

Upstream tips	Protocol tips	Downstream tips
	CHO-K1 cells were cultured in DMEM/F-12 (Invitrogen), and Bosc23 and B16-F1 cells were cultured in DMEM (Invitrogen), all supplemented with 10% fetal calf serum and 100 μg/ml penicillin/streptomycin. Cells were transfected with plasmids encoding secreted ShhN, ShhNp, or sAP-ShhN using PolyFect (Qiagen). Cells were grown for 36 h, washed with phosphate-buffered saline, and incubated in DMEM or serum-free DMEM for various time periods in the presence or absence of stimulators or inhibitors of shedding, followed by ultracentrifugation at 210,000 × g for 60 min to remove cell debris, including membrane-tethered ShhNp.

Protocol tips
CHO-K1 cells were cultured in DMEM/F-12 (Invitrogen), and Bosc23 and B16-F1 cells were cultured in DMEM (Invitrogen), all supplemented with 10% fetal calf serum and 100 μg/ml penicillin/streptomycin. Cells were transfected with plasmids encoding secreted ShhN, ShhNp, or sAP-ShhN using PolyFect (Qiagen). Cells were grown for 36 h, washed with phosphate-buffered saline, and incubated in DMEM or serum-free DMEM for various time periods in the presence or absence of stimulators or inhibitors of shedding, followed by ultracentrifugation at 210,000 × g for 60 min to remove cell debris, including membrane-tethered ShhNp.

Protocol tips

CHO-K1 cells were cultured in DMEM/F-12 (Invitrogen), and Bosc23 and B16-F1 cells were cultured in DMEM (Invitrogen), all supplemented with 10% fetal calf serum and 100 μg/ml penicillin/streptomycin. Cells were transfected with plasmids encoding secreted ShhN, ShhNp, or sAP-ShhN using PolyFect (Qiagen). Cells were grown for 36 h, washed with phosphate-buffered saline, and incubated in DMEM or serum-free DMEM for various time periods in the presence or absence of stimulators or inhibitors of shedding, followed by ultracentrifugation at 210,000 × g for 60 min to remove cell debris, including membrane-tethered ShhNp.

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