CN108421106A - Bioartificial liver system based on people iHep cells and membrane type panel-shaped reactor - Google Patents

Abstract

The invention discloses a kind of bioartificial liver system based on people iHep cells and membrane type panel-shaped reactor comprising the first circulation pump, heparin pump, plasma separator, the second circulation pump and bioreactor, wherein.The import of first circulation pump is connect with blood input pipe, the outlet of first circulation pump is connect with the heparin pump, the plasma separator respectively, and the outlet of the plasma separator is connect with blood efferent duct and the second blood circulation pump intake.The bioartificial liver system of the present invention has sufficiently large capacity to load the liver cell that bodily fuctions can be maintained to need using the bioreactor of artificial liver of biomembrane；The bioreactor of the present invention can be placed on the culture that liver cell is carried out in regular growth incubator；It is long to continue the time used, perfect in shape and function, ensures that the liver cell of culture does not enter blood circulation of human body system to ensure treatment safety.

Description

Bioartificial liver system based on human iHep cells and membrane plate reactor

technical field

The invention relates to the technical field of biological liver, in particular to a biological artificial liver system based on human iHep cells and a membrane-type plate reactor.

Background technique

my country is the country with the most serious liver diseases in the world. Most of the patients are treated through liver transplantation. However, due to the difficulty in obtaining livers, it takes a long time to wait for a liver source, and there is even no chance to get a donor liver. Therefore, most patients have no chance of survival at all. Therefore, an artificial liver system is urgently needed to maintain the patient's life, waiting for the opportunity of liver transplantation, or the opportunity of the recovery of the patient's own liver.

However, due to the high cost of the traditional artificial liver, most patients cannot afford it, the curative effect is very limited, and the mortality rate of liver failure cannot be significantly reduced, so the clinical application is limited. Previous bioartificial liver support systems mainly consisted of hollow fiber reactors filled with a certain amount of hepatocytes or hepatocyte-like cells (such as human hepatocytes, immortalized hepatocytes, hepatoma cell lines, porcine hepatocytes, etc.). There are obvious cell defects, such as difficult cell sources, inability to proliferate on a large scale, poor liver function and safety, etc. In the hollow fiber tube reactor, the liver cells gradually lose their function due to the inability to adhere to the wall. The cells in the reactor may also gradually die due to the influence of toxins. The efficiency of the bioreactor to purify blood decreases with the prolongation of working hours. In order to maintain the curative effect, the reactor with fresh cells needs to be replaced at regular intervals.

To sum up, the existing artificial liver support system cannot completely replace the functions of the whole liver, the detoxification function is monotonous, the plasma consumption is huge, the duration of use is short, and the protein synthesis, fat metabolism, glucose metabolism and detoxification of complete liver cells are not available. Important functions need to be further improved.

Contents of the invention

The purpose of the present invention is to provide a bioartificial liver system based on human iHep cells and a membrane-type plate reactor, which is used to solve the problem of monotonous detoxification function, huge plasma consumption, short duration of use, and imperfect functions of the existing artificial liver support system. and poor safety performance.

To achieve the above object, the present invention provides a bioartificial liver system based on human iHep cells and a membrane plate reactor,

The bioartificial liver system includes a first blood circulation pump, a heparin pump, a plasma separator, a second blood circulation pump and a bioreactor, wherein. The inlet of the first blood circulation pump is connected to the blood input tube, the outlet of the first blood circulation pump is respectively connected to the heparin pump and the plasma separator, and the outlet of the plasma separator is connected to the blood output tube and the blood plasma separator. The inlet of the second blood circulation pump is connected;

The outlet of the second blood circulation pump is connected to the inlet of the bioreactor, and the outlet of the bioreactor is connected to the blood outlet tube.

Preferably, the bioreactor comprises: a housing, a plurality of supports arranged in the housing, a biofilm covering the supports, and a support tube fixedly connected to the supports;

Wherein, the housing is provided with a plasma inlet, a cell culture solution and a gas inlet, the support tube passes through the housing and is fixedly connected with the housing, one end of the support tube is a cell injection port, and the other end For plasma outlet, a plurality of through holes are provided on the side wall of the support tube.

Preferably, the number of the stents is 10-15.

Preferably, the biofilm is a nanofiber film.

Preferably, the housing is made of medical plastic or metal.

Preferably, the housing is cylindrical, and the housing includes an upper cover and a lower cover, and the plasma inlet, cell culture solution and gas inlet are opened on the upper cover.

Preferably, the support is a disc-shaped support, and radial support rods are arranged inside the disc-shaped support, or a plurality of through holes are provided on the disc-shaped support.

Preferably, the bioartificial liver system based on human iHep cells and membrane plate reactors of the present invention also includes a plasma mixer, the plasma mixer has a first inlet, a second inlet and a blood outlet, and the blood outlet is connected to the The blood output tube is connected, the first inlet is connected with the plasma separator, and the second inlet is connected with the bioreactor outlet.

The present invention has the following advantages:

The artificial liver system of the present invention uses a biofilm artificial liver bioreactor with a large enough capacity to hold the liver cells that can maintain the functions of the human body; the bioreactor of the present invention can be placed in a conventional cell culture box for the cultivation of liver cells ; It can be used continuously for a long time, with perfect functions, and ensures that the cultured liver cells do not enter the human blood circulation system to ensure the safety of treatment. It really has the meaning of "artificial liver".

Description of drawings

Figure 1 is a schematic diagram of the composition of the bioartificial liver system based on human iHep cells and membrane plate reactors of the present invention.

Fig. 2 is a structural schematic diagram of the bioreactor of the present invention.

Fig. 3 is a combined structure diagram of the bracket and the support tube of the bioreactor of the present invention.

Fig. 4 is a schematic diagram of the structure of the biofilm of the present invention.

Fig. 5 is a schematic structural view of the stent of the present invention.

Detailed ways

The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention.

As shown in Figure 1, the bioartificial liver system based on human iHep cells and membrane plate reactor of the present invention includes a first blood circulation pump 200, a heparin pump 400, a plasma separator 500, and a second blood circulation pump 300 And the bioreactor 100, wherein. The inlet of the first blood circulation pump 200 is connected with the blood input tube 700, the outlet of the first blood circulation pump 200 is connected with the heparin pump 400 and the plasma separator 500 respectively, and the outlet of the plasma separator 500 is connected with the blood output tube 800 and the second blood The inlet of the circulation pump 300 is connected; the outlet of the second blood circulation pump 300 is connected with the inlet of the bioreactor 100 , and the outlet of the bioreactor 100 is connected with the blood outlet tube 800 . The plasma separator 100 separates blood cells and plasma, and the separated plasma enters the bioreactor 100 through the second blood circulation pump 300, and reacts with the cells in the bioreactor 100. After the reaction is completed, the plasma carries related substances into the blood outlet tube 800, and mixed with the blood cells separated by the plasma separator 500, the blood output tube 800 is connected with the human body, and the substances generated by the reaction are input into the human body, and the blood circulated in the human body is input into the bioartificial liver through the blood input tube system. The bioartificial liver system of the present invention is developed on the basis of the artificial liver support system. It has quite complete liver functions, can completely replace the human liver, and fully possesses the functions of protein synthesis, glucose metabolism, fat metabolism and detoxification of the liver. It can be widely used in the rescue of patients with acute and chronic liver failure, and truly has the meaning of "artificial liver".

Specifically, as shown in Figure 2 and Figure 3, the bioreactor of the present invention comprises: housing 150, a plurality of support 140 that is arranged in housing 150, the biofilm 120 that covers on support 140 and fix with support 140 A connected support tube 110; wherein, the housing 150 is provided with a plasma inlet 131, a cell culture solution and an air inlet 132, the support tube 110 passes through the housing 150 and is fixedly connected with the housing 150, and one end of the support tube 110 is a cell injection port 111 , the other end is a plasma outlet 112 , and a plurality of through holes 113 are provided on the side wall of the support tube 110 . The hepatocytes to be cultured and the culture solution enter the bioreactor 100 through the cell injection port of the support tube 110, and the hepatocytes and the culture solution are distributed to the biofilm 120 through the through holes 113 on the side wall of the support tube 110, and the hepatocytes Reacting on the biofilm 120, the plasma separated by the plasma separator 500 enters the bioreactor 100 through the plasma inlet on the housing 150, and the nutrient solution for cultivating cells enters the bioreactor 100 through the cell culture solution inlet 111, At the same time, air enters the bioreactor 100 through the cell culture solution and the air inlet 132 to provide suitable air, temperature and humidity for the growth of liver cells, and the reacted plasma solution enters the support through the through hole 113 on the side wall of the support tube 110. Then, the reaction liquid flows into the blood output tube 800 through the outlet of the support tube 110, and finally enters the human body.

Wherein, as shown in FIG. 4 , the number of brackets 140 is 10-15, and the surface of each bracket 140 is covered with a biofilm 120 , and the biofilm 120 is a nanofiber film. Hepatocytes can better play the function of hepatocytes when they grow on the wall. The present invention uses a membrane-type plate-shaped bioartificial liver reactor on the bioreactor to make hepatocytes grow well on the biofilm so that The function of liver cells is good, and at the same time, sufficient number of liver cells can be guaranteed to maintain the liver function requirements of the whole human body.

The housing 150 is cylindrical, and the housing includes an upper cover 130 and a lower cover 160, both of which are made of medical plastic or metal. The plasma inlet 131 and the cell culture solution inlet 111 are opened on the upper cover 130 of the casing 150. As shown in Figure 5, each support 140 is a disc-shaped support, and radial support rods 141 are provided in the disc-shaped support 140, or a plurality of through holes are provided on the disc-shaped support 140, and the shape of the through holes can be Regular or irregular shapes such as circles, rectangles, rhombuses, and porous scaffolds can prevent cells from overgrown and prevent plasma flow. At the same time, the growth shape of liver cells can be adjusted according to the shape of the holes and the opening ratio. The shape of the bracket 140 is slightly smaller than the outer diameter of the housing, and the edge of the bracket 140 is in contact with the inner wall of the housing.

The bioartificial liver system based on human iHep cells and membrane plate reactor of the present invention also includes a plasma mixer 600, the plasma mixer 600 has a first inlet, a second inlet and a blood outlet, and the blood outlet is connected to the blood outlet tube 800 , the first inlet is connected to the outlet of the plasma separator 500 , and the second inlet is connected to the outlet of the bioreactor 100 . The plasma reacted by the bioreactor 100 enters the plasma mixer 600 through the reaction liquid outlet of the support tube 110 and the second inlet of the plasma mixer 600, and at the same time, the blood cells separated by the plasma separator 500 pass through the first inlet of the plasma mixer 600. The inlet enters into the plasma mixer 600 , and after being mixed by the plasma mixer 600 , enters the human body through the blood outlet tube 800 .

Furthermore, in the bioartificial liver system of the present invention, a temperature sensor and a pressure sensor can also be installed to sense the temperature and pressure of the plasma entering the output tube, so as to facilitate the control of the pressure and temperature of the plasma output process in the entire bioartificial liver system.

When the bioartificial liver system of the present invention is used, a certain amount of hepatocytes is cultivated by ordinary culture medium method, and the active hepatocytes are harvested. Then, through aseptic operation, an appropriate amount of culture solution containing hepatocytes is injected into the bioreactor 100 through the cell injection port of the support tube, the culture is continued to a certain extent, and it is stored at 0-4°C until use. When the patient who needs the artificial liver system is used, the blood input tube 700 and the blood output tube 800 of the bioartificial liver system are respectively connected to the patient's arterial blood inlet and venous return port for treatment.

The bioartificial liver system based on human iHep cells and a membrane-type plate reactor of the present invention solves the problem of the source of human hepatocytes. The bioreactor is provided with a bracket and a biofilm covering the bracket, which has a large enough capacity Can maintain the liver cells required by human body functions; at the same time, the bioreactor is equipped with a gas exchange channel, so the bioreactor can be placed in a conventional cell incubator for hepatocyte culture, and the hepatocytes grown on the wall have better cell Function: Hepatocytes differentiated from human stem cells are used to maintain the function of the liver. The ingenious design of the reactor ensures that the hepatocytes do not enter the human blood circulation system to ensure the safety of treatment.

Although the present invention has been described in detail with general descriptions and specific examples above, it is obvious to those skilled in the art that some modifications or improvements can be made on the basis of the present invention. Therefore, the modifications or improvements made on the basis of not departing from the spirit of the present invention all belong to the protection scope of the present invention.

Claims (8)

1. a kind of bioartificial liver system based on people iHep cells and membrane type panel-shaped reactor, which is characterized in that

The bioartificial liver system include the first circulation pump, heparin pump, plasma separator, the second circulation pump and Bioreactor, wherein.The import of first circulation pump is connect with blood input pipe, first circulation pump Outlet connect with the heparin pump, the plasma separator respectively, the outlet of the plasma separator and blood efferent duct and The second blood circulation pump intake connection；

The outlet of second circulation pump is connect with the entrance of the bioreactor, the outlet of the bioreactor with The blood efferent duct connection.

2. the bioartificial liver system according to claim 1 based on people iHep cells and membrane type panel-shaped reactor, special Sign is,

The bioreactor includes:Shell, the multiple holders being arranged in shell, the biomembrane being covered on the holder with And the support tube being fixedly connected with the holder；

Wherein, the shell is equipped with plasma inlet, cell culture fluid and gas access, and the support tube passes through the shell And be fixedly connected with the shell, described support tube one end is cell inlet, and the other end is plasma outlet port, in the support tube Side wall be equipped with multiple through-holes.

3. the bioartificial liver system according to claim 2 based on people iHep cells and membrane type panel-shaped reactor, special Sign is,

The quantity of the holder is 10-15.

4. according to claim 2 be based on the bioartificial liver of people iHep cells and membrane type plate pattern shape reactor System, which is characterized in that

The biomembrane is nano-fiber film.

5. the bioartificial liver system according to claim 2 based on people iHep cells and membrane type panel-shaped reactor, special Sign is,

The shell is made of medical plastic or metal material.

6. the bioartificial liver system according to claim 1 based on people iHep cells and membrane type panel-shaped reactor, special Sign is,

The shell is cylindrical shape, and the shell includes upper cover body and lower cover, and the plasma inlet and cell culture fluid are gentle Body entrance is opened in the upper cover body.

7. the bioartificial liver system according to claim 4 based on people iHep cells and membrane type panel-shaped reactor, special Sign is,

The holder is discoid holder, and radial supporting rod or described discoid is equipped in the discoid holder Holder is equipped with multiple through-holes.

8. the bioartificial liver system according to claim 1 based on people iHep cells and membrane type panel-shaped reactor, special Sign is to further include blood plasma mixer, and the blood plasma mixer has the first import, the second import and blood outlet, the blood Liquid outlet is connect with the blood efferent duct, and first import is connect with the plasma separator, second import and institute State bioreactor outlet connection.