Tsingtao amphioxus cell line culture fluid and preparation method thereof
A culture medium and cell line technology, applied in the field of cell biology, can solve problems such as inability to subculture amphioxus cells in vitro
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Embodiment 1
[0038] The specific steps of establishing the Qingdao amphioxus cell line provided by the present invention are as follows:
[0039] 1) Cultivate primary cells from the gills of Qingdao amphioxus: put Qingdao amphioxus in an ice bath until comatose, disinfect the body surface with 70-75% alcohol, dissect aseptically, take the gill tissue of amphioxus, and use cells containing penicillin and streptomycin After rinsing with the culture medium, cut it into small tissue pieces, transfer it to a six-well cell culture plate, spread it evenly, and culture it at a constant temperature at 25-29°C to make it adhere to the wall. Observe the growth status of the cells every day, and add 0.5-1ml cell culture medium; replace 1 / 2 volume of cell culture medium after 3 days. Such as figure 1 As shown, after the primary culture of amphioxus gill tissue, adherent cells and suspension cells can be seen to dissociate from the tissue. The suspension cells are round and the adherent cells are polyg...
Embodiment 2~5
[0045] The culture medium component that is specially used in the above-mentioned method for establishing Qingdao amphioxus cell line provided by the present invention is preferred:
[0046]
[0047]
[0048]
[0049] Preparation Example
[0050] The preparation provided by the invention is specially used to establish the cell culture fluid of described Qingdao amphioxus cell line method:
[0051] Add the components shown in the table into 0.8 liters of ultrapure water (resistivity 18.2Ω), adjust the osmotic pressure to 750-800mOsm / kg, adjust the pH to 7.0-7.2, and use ultrapure water (resistivity 18.2Ω ) Adjust the total volume of the culture solution to 1 liter, mix well, store at 4°C after filter sterilization; add 10% fetal bovine serum before use.
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