ST culture medium and application thereof
A culture medium and vitamin technology, applied in the cultivation process, tissue culture, microorganisms, etc., can solve the problems of easy batch-to-batch differences in cell culture results, exogenous viruses, and unstable sources of serum imports
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Embodiment 1
[0088] In this embodiment, the ST medium used is a serum-free medium, which can be used for the suspension culture of ST cells. The composition and content of the ST medium are shown in Table 1 below:
[0089] Table 1
[0090]
[0091]
[0092] Using the ST medium described in Table 1, the adherent ST cells were gradually and successfully domesticated into suspension ST cells, such as figure 1 shown. The resulting suspended ST cells were treated with the ST medium described in Table 1 at 1x10 6 The density of cells / mL was inoculated in a 125mL shake flask, the working volume was 30mL, and the rotation speed was set at 110rpm. The result: the cell density can reach 1x10 7cells / mL. Under the same experimental conditions, after adding CSFV at a ratio of 0.5% (volume ratio), the cell growth was relatively low, and the cell density reached 0.8×10 7 cells / mL, the expression of the virus was 30% higher than that of the adherent ST cells using the same inoculation strategy. ...
Embodiment 2
[0094] In this embodiment, the ST medium used is a serum-free medium, which can be used for the suspension culture of ST cells. The composition and content of the ST medium are shown in Table 2 below:
[0095] Table 2
[0096]
[0097]
[0098] Using the ST medium described in Table 2, the adherent ST cells were gradually and successfully domesticated into suspension ST cells, such as figure 2 shown. The resulting suspended ST cells were treated with the ST medium described in Table 2 at 0.5x10 5 The density of cells / mL was inoculated in shake flasks, the working volume was 45mL, and the rotation speed was set at 160rpm. After 3 days, the cell density reached 1.8x10 6 cells / mL. Under the same experimental conditions, after adding CSFV at a multiplicity of infection (MOI) of 0.1, the virus expression level was equivalent to that of adherent ST cells.
Embodiment 3
[0100] In this embodiment, the ST medium used is a serum-free medium, which can be used for the suspension culture of ST cells. The composition and content of the ST medium are shown in Table 3 below:
[0101] table 3
[0102]
[0103]
[0104] Using the ST medium described in Table 3, the adherent ST cells were gradually and successfully domesticated into suspension ST cells, such as image 3 shown. The resulting suspension ST cells, using the ST medium described in Table 2, with 5x10 5 The density of cells / mL was inoculated in a 125mL shake flask, the working volume was 30mL, and the rotation speed was set at 150rpm. As a result, the cell density reached 0.8x10 7 cells / mL. Under the same experimental conditions, after adding CSFV strain C according to 0.5MOI, the cell density reached 0.8x10 7 cells / mL, the virus expression reached 6.2±0.3LogTCID50 / mL, which was equivalent to the virus titer of ST adherent cells under the same conditions.
[0105] In summary, using...
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