Mycer immortalized hepatocyte
A liver cell and fusion protein technology, applied in the field of liver cells, can solve the problems of differences between species, unpredictable in vivo toxicity, and low expression of markers
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Embodiment 1
[0046] Materials and methods
[0047] All chemicals were purchased from Sigma Corporation unless otherwise indicated.
[0048] Tissue culture
[0049] All cells were grown in a Millennium incubator (Jencon) at 37°C, 5% CO 2 grow under conditions. Cells were cultured on a regular basis with three medium changes per week. Subculture was performed when 90% confluency was reached.
[0050] culture medium
[0051] Eight media are used during tissue culture. Human fetal liver cells and clones were cultured on all of these media. These media were filter sterilized using 1 liter filter units (Nalgene) and prewarmed to 37°C before use. These eight media are referred to as media A, B, C, D, E, F, G, and H, respectively, and have the following compositions:
[0052] Medium A: arginine-free Williams E standard medium (Gibco), 2.2g / L sodium bicarbonate, 10% dialyzed fetal bovine serum (dFBS) (Gibco 26400-044), 2mM L-glutamine , 100 nM insulin, 100 IU / ml penicillin-streptomycin (Gi...
Embodiment 2
[0099] Materials and methods
[0100] All chemicals were purchased from Sigma (Dorset, UK) unless otherwise indicated.
[0101] Tissue culture
[0102] cells at 37°C, 5% CO 2 grown in a millennium incubator. The medium was changed three times a week, and the cells were passaged when the confluency was about 90%.
[0103] culture medium
[0104] The following kinds of media were used in the culture of liver tissues and clones. The medium was filter-sterilized using a 1 L filter unit (Nalgene, NY, USA) and pre-warmed to 37°C before use.
[0105] Medium A: Dulbecco's modified eagle medium (DMEM) (Invitrogen, Paisley, UK), 10% fetal bovine serum (FBS) (Hyclone Perbio Science, UT, USA), gentamicin (50mg / ml), L - Glutamine (2mM) and Epidermal Growth Factor (EGF) (20ng / ml).
[0106] Medium B: arginine-free Williams E standard medium (Invitrogen, Paisley, UK): DMEM (Invitrogen, Paisley, UK) (1:1), 10% FBS, EGF (20 ng / ml).
[0107] Medium C: DMEM (+Glutamax) (Invitrogen, Paisle...
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