Lung cancer micro-tumor cell model culture method
A technology for lung cancer and tumors, which is applied in the direction of tumor/cancer cells, cell dissociation methods, and cell culture active agents, etc. The effects of rich types, short culture period and high culture stability
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Embodiment 1
[0084] Embodiment 1, preparation is used for cultivating the reagent of lung cancer micro-tumor model
[0085] 1. Sample preservation solution (100mL)
[0086] The specific formulation of the sample preservation solution (100mL) is shown in Table 1.
[0087] Table 1. Sample Preservation Solution (100mL)
[0088]
[0089]
[0090] After the sample preservation solution is prepared, it is divided into 15mL centrifuge tubes, 5mL per tube. After aliquoting, it can be stored at 4°C for 1 month.
[0091] 2. Sample cleaning solution (100mL)
[0092] The specific formula of the sample cleaning solution (100mL) is shown in Table 2.
[0093] Table 2. Sample cleaning solution (100mL)
[0094]
[0095] The sample cleaning solution should be prepared and used immediately.
[0096] 3. Sample dissociation solution (10mL)
[0097] The specific formulation of the sample dissociation solution (10mL) is shown in Table 3.
[0098] Table 3. Sample dissociation solution (10mL)
[...
Embodiment 2
[0162] Embodiment 2, acquisition of postoperative specimen of lung cancer
[0163] 1. Cooperate with tertiary hospitals to obtain samples through the form of clinical research initiated by researchers, and the cooperation has passed the formal medical ethics review.
[0164] 2. The attending doctor selects the patients according to the clinical indications stipulated in the medical guidelines, and selects appropriate samples for in vitro culture according to the clinical indications during the operation. The selection criteria for the samples are: primary lung cancer, pathological stage II or Stage III, the pathological type is non-small cell lung cancer or small cell lung cancer, and the weight of the lung cancer sample exceeds 20mg.
[0165] 3. All enrolled cases are uniformly coded by the date of sample collection + the last four digits of the patient’s hospital number. For example, for a sample provided on January 1, 2020, the patient’s hospital number is T001537474, and t...
Embodiment 3
[0168] Embodiment 3, pre-dissociation treatment of lung cancer tissue samples
[0169] The following operations need to be performed on ice, and the entire operation steps need to be completed within 10 minutes.
[0170] The surgical equipment used in the following operations must be sterilized by high-temperature steam (120°C, 20 minutes) in advance and dried before use.
[0171] 1. After the sample is weighed, clean the surface of the sample with medical alcohol (75% by volume) for 10 to 30 seconds.
[0172] 2. Wash the sample 5 times with sample cleaning solution, and wash the sample 5 times with sterile PBS solution.
[0173] 3. Use ophthalmic scissors, ophthalmic forceps, scalpel and other equipment to carefully peel off the adipose tissue, connective tissue, and necrotic tissue in the sample.
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