Feasibility Study On The In Vitro Organoids Culture Of Lung Cancer Puncture Small Specimen

Objectives Lung cancer is the material cause of cancer-related deaths worldwide,with a5-year overall survival rate of 17%.In order to improve the diagnosis and treatment of non-small cell lung cancer(NSCLC),we studied the CT-guided percutaneous lung biopsy with coaxial puncture needle for histopathological qualitative diagnosis and drug sensitivity test after 3D organoids culture in vitro,screened suitable combination of anticancer drugs for patients,and evaluated the clinical efficacy and feasibility of lung cancer organoid in clinical application.Methods 1 22 patients with suspected malignant lung tumors were scanned with Discovery PET/CT Elite system,and then biopsies were performed with coaxial guided biopsies after locating the location of lung lesions.2 The pathological diagnosis of the lung lesions after puncture was carried out.The remaining puncture tissues were cleaned with tissue cleaning fluid and stored in the tissue preservation fluid.If the pathological findings suggest non-small cell lung cancer,confirm that the case entered the cell culture,and give first-line anti-tumor treatment for the patients according to the latest NCCN,CSCO or Consensus of lung cancer.3 The lung cancer tissues which fit into organoid culture were fully hydrolyzed and dissociated out of lung cancer cells in the enzymolysis solution.4The free lung cancer cells were cultured in three-dimensional medium with specific growth factors and extracellular matrix.The cells were identified as lung cancer cells by microscopic examination,and further expanded and subcultured.5 On 384 well plate,the CellTiter-Glo? reagent was added directly to the lung cancer cells containing serum to determine the survival of the cells on the orifice plate.6 Anti-cancer drug screening,pharmacology and other related drug sensitivity tests were carried out on POLARstar Omega automatic multi-function enzyme labeling instrument for lung cancer organoid cultured in vitro.7 The clinical effect of the first-line anti-cancer regimen was evaluated with the combination of the drugs screened by the drug sensitivity test in vitro organoid culture of lung cancer cells,and the clinical concordance was evaluated.Results 1 22 patients had successful puncture and satisfactory sampling.One patient had a small amount of pneumothorax without catheterization and self-absorption.There were no other complications related to puncture.2 8 of 22 cases had benign pathological changes of lung biopsy tissue.14 cases were malignant tumors,8 cases met the requirements of modeling,and successfully entered 3D organoid culture.After culture,lung tumor cells were identified by microscopic examination.At the same time,2 cycles of clinical empirical anti-tumor therapy were given.3 On 384 well plate,CellTiterGlo?reagent was added directly to the cultured lung cancer cells containing serum to detect cell viability,and all lung cancer organoid were tested for cell viability.The number of cells in each hole detected by the system was 18 to 54.4 According to the first-linetreatment scheme for lung cancer,the drug sensitivity test was carried out on the POLARstar Omega automatic multi-functional enzyme marker after different combination of drugs(including the current clinical treatment drugs of the patients).In 8 cases of lung cancer,drug sensitivity test was positive in 6 cases and negative in 2 cases.5 After 2cycles of treatment in 8 cases of non-small cell lung cancer,the clinical consistency with the results of organoid drug sensitivity test was evaluated: 2 cases of disease progress,2cases of non-small cell lung cancer,2 cases of non-small cell lung cancer.The corresponding drug sensitivity tests were negative.3 patients with stable condition and 3patients with remission were all positive for organoid drug sensitivity test,and the clinical consistency was 100%.Conclusions 1 CT-guided percutaneous lung biopsy can accurately carry out the biopsy of the lung tissue of the lesion,and the characterization of the benign and malignant lesions or the malignant tumor can be confirmed;At the same time,the application of coaxial puncture technology can increase the amount of specimens obtained,and provide adequate tissue samples for related follow-up detection.2 After 3 D organoids culture in vitro,non small cell lung cancer can be sequenced,screened by drug sensitivity test,the combination of anticancer drugs suitable for patients can be selected and provide precise medical treatmentthe,avoided toxicity of drugs and the clinical curative effect can be improved.Figure8;Table7;Reference 66.