Culture Of The Hemangioma Cells And The Analysis On Its Biochemistry And Biology Character

BACKGROUND: Hemangioma, the most common benign tumor of infancy, is belong to the congenital disease. It is caused by the hyperplasia of capillary and vascular endothelial cells(VEC) and the malformation of vascular . It will influence the dissect structure and physiological function of human being, lead to malformation and disfigurement of the visage, even can menace the life. But there aren't any effective treatments at present because the research on its tissue development, differentiation and biological action is nearly lacking.By means of the culture of hemangioma vascular endothelial cells and analysis of its biochemistry and Biology Character, there will be a further understanding about the development, formational condition and disappearance machanism etc. Futhurely the reasonable ways, more effective drugs of treatment will be presented and progress the present situation of clinical treatment. In addition, hemangioma offers a valuable model of human beings for the research on massage machanism about hyperplasia of vascular endothelial cells and development of vascular. It will provide a beneficial clue for the development of the vascular. OBJECTIVE: To master the growth regulation of hemangioma vascular endothelial cells, and to research it on the growth curve, the character of its biochemistry and biology, the difference and similary with the normal adult vascular endothelial cells. It includes the analysis about the existing and expression of the factor â…§ related antigen. To provide the theoretic basis for the clinical treatment of it.METHODS: We us the sterile hemangioma as material to isolate and culture the hemangioma vascular endothelial cells with 0.1% trypsinase to digest the hemangioma VEC, with M199 medium added 10% fetal bovine serum and 75ng/mL FGF, and in the culture box of 5%CO₂,37 C° and saturated humidity. Every two days change the culture liquid. We make them to survive fromgeneration to generation after the hemangioma VEC have grown up to the single layer cells. We identified hemangioma VEC by observing expression of the factor VIH related antigen which only exists in endotheiial cells by the indirect immunofluorescence technique and under a laser scanning confocal electron microscope. The tissue slices of the hemangioma and the normal adult vascular were analyzed about immunohischemistry of the factor W. associated antigen by the laser scanning confocal microscope.RESULTS: Hemangioma cells were cultured successfully, they spreaded on the bottom of cell culture dishes. High-speed growth was found after three weeks. The cells were observed under a phase contrast microscope for cell morphology analysis and appeared as epithelium.The expression of the factor VI associated antigen was analyzed by the indirect immunofluorescence technique, and observed under a laser scanning confocal microscope. We found that the factor VDI associated antigen was positive in the isolated cells and umbilical vein endotheiial cells, negative in A549 lung cancer cells. The results showed that the cells isolated were the vascular endotheiial cells of the hemangioma.Immunohischemistry of the factor Vffi associated antigen in the hemangioma tissue slice showed that vascular in the hemangioma were hyperplastic obviouslyThe tissue slice was observed by the general H.E. staining, the result showed that the cell nucleus express blue, the cell cytoplasm express light red and the red blood cell in the vascular are reddish-orange. The nucleus of the local tissue were denseness, this showed that vascular in the hemangioma were hyperplastic obviously.CONCLUSION: The hemangioma, was benign tumor, its one of the main characteristics is the vascular hyperplastic. The endotheiial cells of vascular and lymph vascular have high hyperplastic, this is the main basic of the vascular tumor.The vascular endothelial cells of the hemangioma can be cultured from generation to generation to establish a cell line. The culture methods have been mastered by our lab. The VEC is similar with the general tumor cell. Contrast with the normal adult VEC, there are difference in the aspects of Morphology, biochemistry and biology.The hemangiomas were mainly caused by the hyperplastic of the vascular endothelial cells. Isolating and culturing the hemangioma VEC successfully will provide experimental material for researching its biological action. The observations of the hemangioma histology might be used to study its histological development. The primary research at level of tissue and cell on its development laid a foundation for its treatment and provided a test model for discovering better clinical drugs.