Human primary tumor cell separation and culture kit
A primary tumor cell, cell culture technology, applied in the direction of tumor/cancer cells, animal cells, vertebrate cells, etc., can solve problems such as cell contamination, achieve high purity, simple and convenient culture effect, and overcome the effect of complex culture process
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Embodiment 1
[0019] see figure 1 , figure 2 , The kit consists of packing box 1, 1 piece of 6-well cell culture plate 2, 5ml cleaning solution 3, 1 bottle of 5ml trypsin digestion solution 4, 1 bottle of 5ml collagenase-hyaluronidase digestion solution 5, 1 bottle of 5ml cell separation Solution 6, 1 bottle of 5ml tumor selective culture solution 7, instruction manual 8.
[0020] The cleaning solution 3 is a sterile RPMI 1640 cell culture solution containing 400 U / mL of penicillin and 400 ug / mL of streptomycin, filled into two 5ml sterile sealed bottles, labeled, and stored at 4°C.
[0021] Trypsin digestion solution 4 is prepared with sterile ultrapure water containing 0.1% trypsin (purchased from Sigma, 100mg), 10g / L polyvinylpyrrolidone trypsin digestion solution, divided into 5ml sterile sealed bottles, and affixed Label and store at 4°C.
[0022] Collagenase-hyaluronidase digestion solution 5 is prepared with RPMI 1640 cell culture medium, wherein the final concentration of collag...
Embodiment 2
[0027] see figure 2 , using this kit: first number each well of the 6-well cell culture plate 2 (a-f).
[0028] (1) After cutting out the tumor specimen, put it into well a of the 6-well cell culture plate 2, and pour cleaning solution 3 to cover the specimen;
[0029] (2) Wash the tumor specimen in well 1, move the specimen into well b, pour cleaning solution 3, and chop or cut the tumor specimen into 1mm 3 size;
[0030] (3) Move the cleaned and shredded tumor specimen block into well c, pour trypsin digestion solution 4, and pre-digest in a cell culture incubator at 37°C for 15 minutes;
[0031](4) After removing from the cell incubator, transfer the pre-digested tumor specimen into well d, pour collagenase-hyaluronidase digestion solution 5, and digest overnight in a refrigerator at 4°C;
[0032] (5) After overnight digestion, collect the digested cell suspension, filter it through a 200-mesh cell strainer into a 50ml centrifuge tube, add cell separation solution 6 to ...
Embodiment 3
[0036] Liver cancer tissue specimens were taken, and the operation steps were the same as those in Example 2 to obtain high-purity liver cancer cells growing in suspension. See results Figure 4 , is the primary liver cancer cells cultured in well f, in the figure: C is the suspended liver cancer cells.
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