In-vitro individualized medicine test method for lung cancer and culture medium

A culture medium and separation culture technology, applied in the field of cell biology and pharmacology, can solve the problems affecting the quality of life of patients, reduce the burden of toxicity and side effects, improve efficiency, and prolong survival time

Inactive Publication Date: 2017-09-12
SHENZHEN RES INST OF WUHAN UNIVERISTY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In addition, pulmonary toxicity caused by antineoplastic drugs, especially chemotherapy drugs, se

Method used

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  • In-vitro individualized medicine test method for lung cancer and culture medium
  • In-vitro individualized medicine test method for lung cancer and culture medium
  • In-vitro individualized medicine test method for lung cancer and culture medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] [Example 1] Medium M for separation, culture, and subculture of human or mammalian primary epithelial cells and primary tumor cells

[0054] Its composition includes: DMEM and Ham's F-12 NUTRIENT MIX mixed medium at a volume ratio of 3:1, while adding 5% fetal bovine serum, 2nM triiodothyronine, 0.5% insulin-transferrin-selenium reagent, 10 uM Y-27632, 10ng / mL epidermal growth factor, 0.4μg / mL hydrocortisone, 1.0nM cholera toxin, 0.5μg / mL amphotericin B, 40μg / mL gentamicin, Activin A 10 ng / ml and 3μg / ml recombinant human R-Spondin-1.

Embodiment 2

[0055] [Embodiment 2] Establishing the operation method of sampling of lung cancer and adjacent tissue samples and its storage and transportation

[0056] 1) With the informed consent of the patient and the patient’s guardian, the lung cancer and adjacent tissue samples used in this experiment are both about 1 cm 3 (Cubic centimeters), taken from a hospital surgical resection sample, the clinical stage of this patient is poorly differentiated lung adenocarcinoma, no other treatment.

[0057] 2) Under clean room conditions, in strict accordance with the standard of aseptic operation, the surgical samples are collected and stored in test tubes, and the sample collection fluid is added according to the sample size, generally 3-5mL, the sample collection fluid contains 2% penicillin / streptomycin And 100ug / ml nystatin PBS, because the sample size is still sufficient, it is divided into two at the same time to try direct isolation culture and long-term liquid nitrogen storage.

[0058] 3) ...

Embodiment 3

[0060] [Example 3] A method for establishing the primary isolation and culture of lung epithelial cells and tumor cells of lung cancer patients with rapid proliferation in a short period of time includes the following specific steps:

[0061] 1) Before the sample is directly separated and cultivated, the ultra-clean workbench is turned on and the UV lamp is turned on. After 30 minutes of work, turn off the UV lamp, turn on the fan to adjust the air volume, and ensure that the wind speed in the working area is always in the ideal state. Start experiment

[0062] 2) Prepare 5 100mm sterile cell culture dishes marked as ①②③④⑤ and place them in order. Add 10 mL of absolute ethanol (analytical purity, flammable, and keep away from the source of ignition) to the petri dish ②, and the petri dishes ③ and ④ respectively Add 10 mL of PBS buffer in an ice bath.

[0063] 3) Preparation of digestion solution: the above medium M containing 0.2mg / mL collagenase / hyaluronidase (07912, STEMCELL) (mix...

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Abstract

The invention discloses an in-vitro individualized medicine test method for lung cancer and a culture medium, and belongs to the field of cell biology and pharmacology. The in-vitro individualized medicine test method and the culture medium have the advantages that the culture medium M can be applied to normal epithelial cells and primary tumor cells of human or mammals, accordingly, in-vitro culture systems for tumor cells and para-carcinoma cells of lung cancer patients can be established, and primary cells, with continuous passage and quick proliferation functions, of patient individuals can be obtained; the culture systems can be used for detecting the sensitivity and the safety of medicines or combination groups of the medicines, and accordingly stable, accurate and reliable lung cancer individualized treatment-medicine sensitivity test standard detection process systems can be established; individualized sensitive chemotherapy medicines or compositions screened by the aid of the lung cancer individualized treatment-medicine sensitivity test standard detection process systems can be combined with clinical medicine or relevant subjects, accordingly, clinical individualized treatment schemes can be formulated, and the in-vitro individualized medicine test method and the culture medium can ultimately serve for clinical application.

Description

Technical field [0001] The invention belongs to the field of cell biology and pharmacology, and relates to the establishment and culture medium of an in vitro individualized drug testing system for lung cancer. Background technique [0002] Lung cancer is the most common malignant tumor with the highest mortality rate in China and the world, and it is the number one killer of human health. Lung cancer can be divided into non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC) according to pathological types, accounting for about 80% and 20% of all lung cancers. In non-small cell lung cancer, squamous cell carcinoma accounts for about 20%-35%, and adenocarcinoma accounts for about 30%-45%. The late staging of most patients at the time of diagnosis of lung cancer is an important reason that affects the prognosis of lung cancer. Early stage lung cancer can achieve a better prognosis through multidisciplinary comprehensive treatment and can obtain better clinical benefi...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12N5/09C12Q1/02
CPCC12N5/0625C12N5/0688C12N5/0693C12N2500/25C12N2500/84C12N2501/01C12N2501/04C12N2501/11C12N2501/16C12N2501/415C12N2509/00G01N33/5011G01N2500/10
Inventor 李晖陈雨叶立娜吴小婷赵天龙张康蔡铠红魏高斌曾志宏
Owner SHENZHEN RES INST OF WUHAN UNIVERISTY
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