Method for culturing porcine mammary epithelial cells by 3D
A mammary epithelial cell, 3D technology, applied in the field of 3D culturing porcine mammary epithelial cells, can solve the problem that porcine mammary epithelial cells have not been reported yet, and achieve the effect of time-saving and simple operation.
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Embodiment 1
[0028] The present embodiment provides a method for culturing porcine mammary epithelial cells in 3D, comprising the following steps:
[0029] (1) Preheat the 24-well plate at 37°C;
[0030] (2) Digest pig mammary gland epithelial cells before generation 10 into single cells with trypsin and centrifuge at 1000rpm for 4min, discard the supernatant, and keep the precipitate;
[0031] (3) Resuspend the precipitate in a mixed solution prepared at 5°C with Matrigel and Advanced DMEM / F12 complete medium at a volume ratio of 1:1, and adjust the concentration of porcine mammary gland epithelial cells to 4×10 4 cell / ml;
[0032] (4) Drop the resuspension solution into the 24-well plate according to the amount of 30-40ul per drop, drop one drop into each well, incubate the 24-well plate at 37°C for 40min to form a gel;
[0033] (5) Add 550ul Advanced DMEM / F12 complete medium to the 24-well plate and continue culturing for 2 days, and replace the Advanced DMEM / F12 complete medium every...
Embodiment 2
[0037] The present embodiment provides a method for culturing porcine mammary epithelial cells in 3D, comprising the following steps:
[0038] (1) Preheat the 6-well plate at 37°C;
[0039] (2) Digest pig mammary gland epithelial cells before generation 10 into single cells with trypsin and centrifuge at 1200rpm for 4min, discard the supernatant, and keep the precipitate;
[0040] (3) Resuspend the precipitate in the mixed solution prepared by Matrigel and Advanced DMEM / F12 complete medium at a volume ratio of 1:1 at 6°C, and adjust the concentration of porcine mammary gland epithelial cells to 1×10 4 cell / ml;
[0041](4) Drop the resuspension solution into a 6-well plate according to the amount of 30-40ul per drop, drop 6 drops into each well, incubate the 6-well plate at 37°C for 40 minutes to form a gel;
[0042] (5) Add 2 mL of Advanced DMEM / F12 complete medium to the 6-well plate to continue culturing for 3 days, and replace the Advanced DMEM / F12 complete medium every o...
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