The Research Of Cell Culture-method And The Outside-pleural-membrane Animal Model Establishment Of Human Malignant Pleural Mesothelioma

Objective:To used three cell culture methods to raise the original generation cells of the human Malignant Pleural Masothelioma(MPM) and discussed their biology characteristics,and also compared three methods to find whether they had some differences.To established mouse transplanted model of malignant mesothelioma and analyzed its pathology characteristics.Hope it could lay the foundations to the clinical research and the Pharmacology of the human malignant pleural mesothelioma.Methods:The specimens came from patients who visited the thoracic surgery department of the first affiliated hospital of Kunming Medical College.They all were diagnosed malignant pleural membrane mesothelioma by pathology.We obtained the hydrothorax specimens and the pleural membrane specimens by puncture.The tumour cells were drown from the plural fluid by centrifugalization and they were cultivated the original generations.The experiments used RPMI-1640 nutrient fluid,DMEM nutrient fluid and DMEM/F-12(1:1) nutrient fluid which all contained 10%embryo cow blood serum.Observed the cell morphous through light microscope and inverted microscope.Drew the cell growth curve,computed the cell's doubling time and fission index.Established mouse hypodermic transplanted tumor model and observed the transplanted tumor cells' ultrastructures under the electron microscope.After the convention pathological section HE dyeing,observed the transplanted tumor cells' pathology characteristics.Results:1.To successfully got the human malignant pleural membrane mesothelioma cell from the pleural fluid.It was raised well and transferred stably in the DMEM and DMEM/F-12(1:1) complete nutrient fluid.From the pleural membrane specimen,we didn't get the turnour cell.2.The tumor cells from the pleural fluid had the same cytology and pathology characteristics with the human malignant pleural membrane mesothelioma.The cells showed short spindle-shape,kind of polygon or anomalous shape.The nuclear-cytoplasmic ratio was bigger.The cells' ability of pasting to wall was weak,and ability of overlapping growth was more obvious.3.The mouse hypodermic transplanted tumor cells had same pathology characteristics with the human malignant pleural membrane mesothelioma cells. The tumor cells were big and showed anomalous shape.The cytoplasm were plentiful and acidophilia.The nuclear was big and anomalous,and the nuclear fission happened frequently.Conclusions:1.This test successfully accomplished the human malignant pleural membrane mesothelioma cells' original generation raise.2.The DMEM/F-12(1:1)complete nutrient fluid is the best culture medium which used to raise the human malignant membrane mesothelioma cells from the pleural fluid.3.Successfully established the mouse hypodermic transplanted model of human malignant pleural membrane mesothelioma cells.4.The human malignant pleural membrane mesothelioma cell from the mouse hypodermic transplanted model had same pathology characteristics with the human malignant pleural membrane mesothelioma cell.