Isolation Of Hepatic Stellate Cells And The Impact Of Cell Injury On The Cell Morphology And Cell Polarity

Hepatic fibrosis is the excessively collagen accumulation during the tissue repair response to chronic liver injury due to the novel synthesis. The cells and soluble factors participating in this response in the liver are the hepatic stellate cells. Hepatic stellate cells are non-parenchymal, quiescent cells in normal, whose main functions are to store vitamin A and probably to maintain the normal basement membrane type matrix. However, numerous References indicate that in response to liver injury stellate cells undergo an "activation" process in which they lose vitamin A, become highly proliferative, and synthesize 'fibrotic' matrix rich in type I collagen.HSCs were isolated by PERCOLL density gradient centrifugation after the hepatocytes obtained from SD adult rats which were digested with pronase and collagenase infused via portal vein. The cell viability was determined by trypan blue exclusion test. The purity of HSCs was identified by detectingα-smooth muscle actin immunohistochemical staining. This is the foundation for the next experiments.Myofibroblast were isolated by using specific approach that foreign protein have been implanted into New Zealand rabbits'cavitas abdominalis for a period of time. The cells are planted in coverslip with high density and studied the form and orientation after scratch.In the final analysis tissue injury is cell injury which might cause by two kinds of pathological phenomena: First, tissues will lose their functions. Second, the gap after cell death will change the structure of the tissue. From the view of cellular mechanics, the gap after cell death can also be called the destruction of cells stressed. Then the cells will get a signal and respond. In this research, we make mechanical damage on cells in vitro and use bio-mechanics, cell biology, immunohistochemistry and computer image processing to study cell's shape and polarity effects. SPSS was used for statistics analyse.The major founding are as follow:①HSCs were isolated by PERCOLL density gradient centrifugation after the hepatocytes obtained from SD adult rats which were digested with pronase and collagenase infused via portal vein. The cell viability was determined by trypan blue exclusion test. The purity of HSCs was identified by detectingα-smooth muscle actin immunohistochemical staining. ②Static relaxation was exerted on Myofibroblast from processed New Zealand rabbits(1.5kg) using specific approach that foreign protein have been implanted into rabbits'cavitas abdominalis for a period of time. The cells are planted in coverslip with high density and cultivated for 60h.③Scratch 3 days, 5 days, 10 days of the normalized cultured myofibroblast, continuous observation of cell shape and orientation changes, polarity effects.④Statistical analysis of cell characterization angle, length, etc., Get the conclusion of effects by cutting the different number of days.⑤Through the analysis of the angle of MFB with the line of scratching, we get conclusion that there is a positive regulatory effect to MFB by mechanical damage.