Preliminary Study Of Human Colorectal Cancer Cells To Build Plants

Objective Colorectal cancer is a very common digestive system cancer.At present the fundamental research is focused on studying its pathogenesis,and clinical research is centralized in its early diagnosis and therapy.With the increasing recognition of this disease,researchers gradually realize that both fundamental research,clinical research and prevention need thoroughly understanding of biological characteristics colorectal cancer cell.Establishment of cell line provides a useful target material to study cell biological characteristics.The present experiment plans to establish human colorectal cancer cell line for better understanding of colorectal cancer cell biological characteristic and provide a useful cell modal for studying colorectal cancer pathogenesis mechanism.Methods This experiment consists of four parts.The first part is in vitro culture tumor tissue. Tissue was obtained from a surgically resected fresh colorectal cancer,moderately differentiated tubular adenocarcinoma gradeâ…¡,occurring in a 70-year-old male.Using tissue piece andâ…£collagenase digestion do primary cell culture.The cells were proliferated and passaged in high glucose DMEM medium,supplemented with 10%heat-inactivated FBS then observation cell morphours characteristic and cell growth condition in inverted microscope.The second part is identification cultured cells,including histological stain(HE)to observe cell shape and to see whether or not have tumor morphology characteristic;Using transmission electron microscope to view cells ultramicrostructure,and from cells inside to see if they accord with cancer cells; Using immunocytochemistry identify cytokeratin to see if it fits to cancer is origining from epithelial tissue characteristic;The third part is to describe cultured ceils biological characteristic. Applying MTT assay to draw cell growth curve,to detect cell DNA index number and analysis cell ploidy;observation cell invasion by boyden chamber technique;the four parts is animal experiment,viewing cell oncogenicity by subcutaneous inoculation on BALB/C nunu node mice back;4 weeks later put node mice to death and obtain tumor tissue from node mice to make pathological diagnosis and HE stain.Results(1)Colorectal cancer cell was successfully cultured in vitro for 4 months The cell culture got initial successs.(2)The cell growth curve showed,when doing primary culture cell grown slowly,when passaging cell grown faster,one week one passage.(3)The cells' morphous were fusiform and irregular polygon,having aggregation growth condency.HE stain showed karyomegaly and blue-stain,according with cancer cell morphology characteristic.Immunohistochemistry showed cytoplasm yellow-stain CK positive. TEM showed big cell nuclear,abundance in mitochondria,cell surface microvillus and cell-cell junctional complex,according with metabolism vigorous cancer cell characteristic.Health human cell is diploid cell,DNA index result showed heteroploid.(4)Animal experiment.Three days later after appeared small tumor transplant,a week later the tumor transplant begins to grow big.4 weeks later,node mice had a bad appetite, weight and body surface temperature were lowly,and dislocation put mice to death.HE stain showed cancer cell morphology characteristics.Conclusion This experiment successfully does primary cell culture,stable passage description and identification cell biological character,testifing the cell origining from epithelial tissue,having cancer cell's main morphological character and have oncogenicity and invasion characteristics,give a basement for future research and successful establishment cell line.