Liver perfusion method for separating rat liver cells and perfusion apparatus of method

Abstract

The invention belongs to the technical field of animal liver perfusion for separation of liver cells and in particular relates to a liver perfusion method for separating rat liver cells and a perfusion apparatus of the method. In the method, according to a principle design of a classic two-step perfusion method, weight changes of rat livers are monitored in real time in a second step of enzyme liquid perfusion, and stop of the perfusion process is automatically controlled at the right time. The perfusion apparatus comprises a weighing platform, a perfusion system and a data processing system, wherein the perfusion system comprises a buffer solution tank, a peristaltic pump, a thermostat, a bubble trap and a puncture needle connected in sequence by virtue of infusion tubes; the weighing platform is electrically connected with the data processing system; rat livers are arranged on the weighing platform. The invention provides a perfusion apparatus capable of continuously monitoring liver weight changes in the perfusion process, and perfusion can be automatically stopped at the right time, so that viability and quantities of liver cells separated after liver perfusion are obviously improved, and the perfusion apparatus has significant substantive features.

Description

technical field [0001] The invention relates to the technical field of animal liver perfusion separation of hepatocytes, in particular to liver perfusion and a perfusion device for rat hepatocyte separation. Background technique [0002] Seglen published a two-step perfusion protocol for rat hepatocyte isolation in 1976. In the first step of the two-step perfusion, the Ca-free 2+ Buffer introduced into the liver to remove Ca from cell junctions 2+ . This step is to prepare for the second step of perfusion. Once the cellular connections are broken, an enzymatic buffer is introduced to the liver to break down the extracellular matrix. As a result, hepatocytes will be released from cell junctions and extracellular matrix. At the final stage, an isolated hepatocyte suspension can be obtained by gently mechanically agitating the liver to disperse the hepatocytes into the medium. However, the results of two-step perfusion are highly sensitive to the duration of enzyme buffer...

AI Technical Summary

Problems solved by technology

However, the real-time monitoring of organ perfusion equipment at home and abroad focuses more on maintaining the perfusion pressure, temperature, flow rate, pH value of the buffer, etc. The purpose is to maintain the activity of the isolated organ, and the purpose of the two-step perfusion method is to separate cells from the organ. , so the monitoring system developed for the perfusion of these organs is not suitable for the two-step perfusion method

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