Rat Hepatic Parenchymal Cell Isolation and Culture Kit

Matters Need Attention

1. In this experiment, you need to prepare PBS buffer, trypan blue staining solution, syringes, scissors, forceps, culture vessels, venous indwelling needle, and other reagents and consumables. If the cultivation needs to be expanded, please prepare complete medium independently.
2. If the isolation process of primary hepatocytes is not handled properly, contamination can occur easily. Ensure that all equipment used in the experiment is sterile.
3. Primary hepatocytes cannot be passaged. Do not use various digestive enzymes to digest and passage the fully confluent hepatocytes after extraction.
4. Hepatocytes are sensitive to fluid shear forces. During the isolation process, avoid vortexing or excessive pipetting when resuspend the cell pallet, in order not to affect cell viability.
5. Liver tissue perfusion can be performed by peristaltic pumps, infusion machines and manual operation. During the process, pay attention to controlling the perfusion flow rate to avoid incomplete or excessive tissue digestion. The following steps take peristaltic pumps and manual perfusion as examples, and the perfusion method can be adjusted according to specific laboratory conditions.
6. The operation of rat liver perfusion digestion is complicated. Prior to formal experiments, it is recommended to practice pre-perfusion to familiarize yourself with operational procedures and avoid unnecessary waste.
7. Due to the bloody process of rat perfusion, this manual does not provide actual photos for reference, but only schematic diagram. If necessary, please contact the technical support to obtain actual photos of the experimental process.

Kit Components