Rat Hepatic Parenchymal Cell Isolation and Culture Kit

(1) This experiment requires the following reagents and consumables: PBS buffer, trypan blue staining solution, syringes, scissors, forceps, culture dishes, intravenous catheters, etc. If you need to expand the culture, please prepare complete medium on your own; (2) During the isolation of primary hepatic parenchymal cells, contamination can easily occur if the procedure is not performed carefully. Please ensure that all experimental tools are sterile; (3) Primary hepatic parenchymal cells cannot be passaged. Do not use various digestive enzymes to digest and passage the hepatic parenchymal cells after extraction; (4) Hepatic parenchymal cells are sensitive to fluid shear stress. During the separation process, do not use vortexing, excessive blowing, or other methods to resuspend cell precipitates, as this may affect cell viability; (5) Liver tissue perfusion can be performed using methods such as peristaltic pumps, perfusion machines, or manual operations. During the process, it is important to control the perfusion flow rate to avoid incomplete or excessive digestion of the tissue; (6) Rat liver perfusion digestion is a complex procedure. It is recommended to perform a perfusion practice run before using this kit to master the technique of liver perfusion before using the kit, to avoid unnecessary waste. This kit is based on an improved classic in situ perfusion digestion method for the separation and extraction of rat hepatic parenchymal cells. After perfusion digestion, separation and purification of liver tissue from one rat, each test (1T) can yield more than 2 × 10? hepatic parenchymal cells. Immunofluorescence identification shows that the cell purity (CK-18 positivity rate) is >90%.