CN201286882Y - Perfusion-type bioreactor of multi-layer membrane structure - Google Patents

Abstract

The utility model provides a perfusion bioreactor with a multi-layer diaphragm structure for a biological artificial liver. The bioreactor is composed of a cylindrical box, a liquid outlet, a liquid inlet, a concentric circular diaphragm and a solid circular diaphragm. There are grid-shaped pores, the diaphragm is fitted in the box through the groove, and the liquid inlet and liquid outlet are respectively equipped with screw caps. The utility model provides a three-dimensional growth environment for the cells, and the cells are evenly distributed, so that the cells can maintain active functions for a long time; the blood flows through the multi-layer membrane structure in the reactor, and directly contacts with the cells, which can fully exchange substances; the utility model design Reasonable, compact structure, easy to enlarge, and convenient to transport.

Description

Multilayer diaphragm structure perfusion bioreactor

technical field

The utility model belongs to the technical field of biomedicine, and relates to a medical device, which is a core device for providing treatment for a biological or mixed artificial liver—a perfusion bioreactor with a multilayer diaphragm structure for an artificial liver.

Background technique

Bioartificial liver is generally composed of cell components and extracorporeal circulation device (bioreactor), which can temporarily replace the liver function of severe disease, so that patients can successfully transition to liver transplantation or recover spontaneously. It is a necessary supplement for liver transplantation treatment. Applications in the treatment of failure are promising. At present, research centers in many countries in the world have conducted in-depth research on cells and bioreactors, and some bioartificial liver systems have entered the clinical verification stage, but the effect is not satisfactory, and there is no established bioartificial liver system in China. Liver products.

The bioreactor is the core part of the bioartificial liver support system in vitro, and its performance has a direct impact on the function of hepatocytes cultured in vitro, and is further related to the efficacy and safety of the bioartificial liver support system. The current bioreactors can be roughly divided into: hollow fiber bioreactors, flat monolayer bioreactors, perfusion bed or scaffold bioreactors, microcapsule suspension bioreactors and other types. Among them, the bioartificial liver system developed around the hollow fiber technology is widely used. However, the distribution of liver cells in the hollow fiber tube is uneven, and the semi-permeable membrane barrier or collagen layer is not conducive to sufficient material exchange between the cultured cells and the patient's blood. Recently, with the improvement of liver cell in vitro culture technology and the research and development of new biopolymer materials, high-tech technologies such as photoetching and microfluidics have also been gradually applied to the development of reactors, and the construction of bioartificial liver reactors has also been continuously improved. But in general, a successful bioreactor should meet the following conditions: 1. Provide sufficient space for cell growth to support about 10% of liver function in a patient with liver failure, equivalent to about 15 billion liver cells. 2. Good biocompatibility, which can effectively maintain the activity of liver cells, especially the functional activity. The research on polymer porous scaffolds and membrane materials has made great progress. In addition, factors including appropriate oxygen and nutrient supply, establishment of metabolite excretion channels, and protection of cells from physical, chemical, and host immune attacks should all be considered. 3. Easy to enlarge, convenient to transport, routinely sterilizable and resistant to low temperature storage, etc. In short, trying to simulate the growth microenvironment of hepatocytes in vivo and maximizing the function of mature hepatocytes in vivo is the principle and trend in the design of various reactors today.

Contents of the invention

The utility model simulates the physiological structure of the human liver lobule, and based on the new membrane material, provides a multi-layer diaphragm structure perfusion bioreactor for artificial liver, that is, a biological artificial liver support system provides a multi-layer membrane structure perfusion type Bioreactor. The bioreactor is composed of a cylindrical box, a liquid outlet, a liquid inlet, a concentric circular diaphragm and a solid circular diaphragm. There are grid-shaped pores on the edge, and the concentric circular diaphragm and the solid circular diaphragm are fitted in the box through grooves. The liquid inlet and liquid outlet are connected with screw caps respectively, and the screw caps are made of 600-mesh cell mesh material. The cabinet is made of transparent plexiglass.

Polymer membrane materials such as polyethersulfone membrane can be selected for concentric circle diaphragm and solid circle.

The perfusion bioreactor with multi-layer membrane structure for artificial liver provided by the utility model has the following advantages: (1) The new membrane material provides a three-dimensional growth environment for cells, so that the cells can maintain active functions for a long time. (2) Compared with the porous scaffold, the cells are evenly distributed and the microenvironment is consistent. (3) The blood flows through the multi-layer membrane structure in the reactor and directly contacts with the cells, which can fully exchange substances. (4) The design is reasonable, the structure is compact, easy to enlarge, and convenient to transport.

Description of drawings

Figure 1: Schematic diagram of the perfusion bioreactor with multi-layer membrane structure for artificial liver.

Figure 2: Schematic diagram of the longitudinal section of a perfusion bioreactor with multi-layer membrane structure for artificial liver.

FIG. 3 : Schematic diagram of the concentric diaphragm 4 .

Fig. 4: Schematic diagram of the solid circular diaphragm 5.

Figure 5: Schematic diagram of the use of the bioartificial liver perfusion system.

Detailed ways

The utility model is further described in conjunction with the accompanying drawings and embodiments.

Example 1

See Fig. 1-4, the perfusion bioreactor with multi-layer diaphragm structure for artificial liver, the reactor is composed of cylindrical box 1, liquid outlet 2, liquid inlet 3, concentric circular diaphragm 4 and solid circular diaphragm 5 , the inner wall of the box body 1 is provided with a groove 6, the concentric circular diaphragm 4 is provided with a channel 7, and the channel 7 is a passage for liquid to flow through, and the edge of the solid circular diaphragm 5 is provided with a grid-like hole 8, which facilitates The liquid overflows, and the concentric circular diaphragm 4 and the solid circular diaphragm 5 are fitted in the box body 1 through the groove 6 to form a sufficient material exchange environment with a multi-layer film structure. The liquid inlet 3 and the liquid outlet 2 are connected with a screw cap 9 and the screw cap 10 are connected by threads, and the screw cap 9 and the screw cap 10 are prepared from 600-mesh cell mesh material. Casing 1 selects transparent plexiglass for use.

The concentric circular diaphragm 4 and the solid circle 5 can be made of polymer membrane materials such as polyethersulfone membranes, which can provide a three-dimensional growth environment for exogenous hepatocytes, which is closer to the growth conditions in vivo, and make the cell functions more perfect.

Example 2

Referring to Fig. 5, the use process is as follows: the blood 11 flows out, and is separated into cell components and plasma components through the plasma separator 12, wherein the plasma components enter the extracorporeal circulation pool 13, then enter the reactor 14 from the liquid inlet 3, pass through the solid circular diaphragm 5 edge mesh hole 8 overflows, and then continues to overflow from the central channel 7 of the concentric circular diaphragm 4, the overall flow of plasma is S-shaped, and the hepatocytes grown on the concentric circular diaphragm 4 and the solid circle 5 are in the flow process Direct contact (hepatocytes are pre-inoculated, see below for specific steps), after sufficient substance exchange, the liquid flows out through the liquid outlet 2 and returns to the circulation pool 13, and after multiple cycles, it is reinfused into the body through the pipeline 15 together with the cell components.

Exogenous hepatocytes can be selected from primary human, porcine hepatocytes or immortalized hepatocyte strains, and the pre-inoculation steps of hepatocytes are as follows:

1. Take hepatocytes in the logarithmic growth phase, digest them with 0.25% trypsin-0.05% EDTA, and prepare a cell suspension with a concentration of 1-5×10*7/ml in high-glucose DMEM medium containing 10% fetal bovine serum.

2. Open the cell screen screw cap 10 of the liquid outlet 2, inject the cell suspension into the reactor 14, and the cells enter the reactor box 1 and attach to the concentric circular diaphragm 4 and the solid circular diaphragm 5 of the polymer material It grows three-dimensionally, and after about 24 hours, it is firmly attached, and the bioartificial liver perfusion system can be formed as shown in Figure 5.

Claims (4)

1. a multilayer diaphragm structure perfusion type bioreactor is characterized in that: the bioreactor consists of a cylindrical casing (1), a liquid outlet (2), a liquid inlet (3), a concentric circle diaphragm ( 4) and a solid circular diaphragm (5), the inner wall of the box (1) has a groove (6), the center of the concentric circular diaphragm (4) is provided with a channel (7), and the edge of the solid circular diaphragm (5) is provided with Grid-shaped pores (8), concentric circular diaphragms (4) and solid circular diaphragms (5) are fitted in the box (1) through grooves (6), liquid inlets (3) and liquid outlets (2) A screw cap (9) and a screw cap (10) are respectively provided for threaded connection. 2. A perfusion bioreactor with a multilayer diaphragm structure according to claim 1, characterized in that: the concentric circular diaphragm (4) and the solid circular diaphragm (5) are made of polymer membrane materials. 3. A perfusion bioreactor with multi-layer membrane structure according to claim 1, characterized in that: the casing (1) is made of transparent plexiglass. 4. A perfusion bioreactor with a multi-layer membrane structure according to claim 1, characterized in that: the screw caps (9), (10) are made of 600-mesh cell mesh material.

Images