A three-dimensional culture method of hepatocytes in vitro

Abstract

The invention discloses a cell sphere gathering culture medium. The cell sphere gathering culture medium comprises a culture area and a fence connected to the periphery of the culture area. The culture area is provided with multiple sphere gathering holes, wherein the lower half portions of the sphere gathering holes are spherical-segment-shaped cavities. The invention further discloses a medium die which comprises a first platy die plate and a second platy die plate, wherein the first platy die plate and the second platy die plate are mutually nested to form a cell sphere gathering culture medium pouring forming cavity. The first die plate is internally provided with a boss and a groove formed adjacent to the boss in the circumferential direction. Multiple protruding points are arranged on the surface of the boss, wherein the upper half portions of the protruding points are spherical segment bodies. A through hole matched with the outer contour of the outer side groove wall of the groove is formed in the second die plate. The invention further discloses a method for conducting animal cell in-vitro three-dimensional sphere gathering culture by utilizing the cell sphere gathering culture medium. The in-vitro three-dimensional culture method for the hepatic cell is relatively low in cost, controllable in scale and simple in process, and the culture effect is greatly superior to that of two-dimensional plate culture.

Description

technical field [0001] The invention relates to an in vitro culture technology of animal cells, in particular to an in vitro three-dimensional culture method of hepatocytes. Background technique [0002] As a liver substitute, hepatocytes are widely used in drug screening, bioartificial liver, liver transplantation research and other fields. In the traditional in vitro cell plate two-dimensional culture, the surface of the hepatocyte growth culture vessel, the cells grow in an adherent island shape, and the cell protein, urea and drug metabolism functions are not easy to maintain, which is very different from the hepatocytes in the body, and cannot replace the hepatocytes in the body Complete liver function, in order to better simulate the physiological conditions of liver cells in vivo, increase the interaction between cells and cells and extracellular matrix, in order to enhance the function of cultured liver cells in vitro, and restore the polarity of liver cells, has dev...

AI Technical Summary

Problems solved by technology

Among them, the suspension spinner bottle culture takes a long time for the cells to form spheres and the rate of spheres is low, and the obtained cell spheres have different sizes and apertures. At the same time, the instrument cannot accurately simulate the physiological state of the cells during the shaking process; by adding polymer substances, the cells can reach Aggregation state, but there are gaps between cells, so that there are defects in the communication between cells. At the same time, if the diameter of the obtained cell sphere is too large, there will be cell necrosis in the center of the cell sphere; the use of photoresist technology is high, and the price Expensive, complicated process, etc.

Images