Tumor sample pretreatment method
A sample and tumor technology, applied in the field of pretreatment of tumor samples, can solve the problems of culture failure, microbial contamination, and low number of clones, and achieve the effect of improving the success rate of culture and reducing the probability of microbial contamination.
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Embodiment 1
[0035] In this protocol, the inventors cultured a total of 34 tumor samples, of which the first 9 samples used the conventional sample processing method, and the other 26 samples used the pretreatment method of the present invention.
[0036] The specific method of common pretreatment: After the sample is obtained, put it into the transfer buffer, transport the sample to the laboratory at 4-8°C in the sample transfer box, and then wash it with HBSS buffer + 1% double antibody for 5 times, each time for 5 minutes, and then use Cut into 1-2mm tissue pieces with scissors, add 5ml of enzymatic solution for treatment, and repeat the digestion several times until the tissue is completely digested, then centrifuge the collected cell fluid, wash 3 times with HBSS buffer, count, seed the plate, and matrix After the gel was solidified, the corresponding medium was added for cultivation.
[0037] The pretreatment method of the present invention: after the sample is obtained, put it into ...
Embodiment 2
[0041] In this protocol, samples from patients with lung squamous cell carcinoma were used, and the transfer buffer proposed by the present invention was used to store them at 4°C for 4 days, and then cultured. The result is as figure 2 As shown, the results show that the transfer buffer proposed by the present invention can effectively maintain the activity of the sample for up to 4 days at 4°C, and can still successfully culture organoids.
Embodiment 3
[0043] In this program, 10 samples were selected and treated with washing buffer in different sequences.
[0044] 10 cases in the control group:
[0045] 1) Perform the second washing treatment on the fresh tumor samples in the washing buffer, that is, wash the fresh tumor samples in the washing buffer 1 for 5 or 6 times, each time for 4-6 minutes; Wash in Washing Buffer 2 for 2-4 minutes; wash the tissue washed in Washing Buffer 2 in Washing Buffer 3 for 2-4 minutes; wash the tissue washed in Washing Buffer 3 in Washing Buffer 4 at 4 Soaking for 28-32 minutes under the condition of ℃; and washing the tissue soaked in the washing buffer 4 in the washing buffer 5 for 5 or 6 times, each time for 4-6 minutes.
[0046] 2) Excluding the samples after the first cleaning treatment, so as to remove fat, blood, necrosis and tissues with a content of interstitium greater than 20%;
[0047] 3) Perform the first cleaning treatment on the removed tissue. Wash the tissue samples in washi...
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