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Cell reactor and artificial liver support system comprising same

A cell reactor and cell technology, applied in the field of biomedicine, can solve the problems of difficult to observe the growth of liver cells, poor growth of liver cells, etc. Effect

Active Publication Date: 2010-06-16
苏州瑞徕生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It can also solve the problem of poor growth of hepatocytes directly cultured in a bioreactor and difficulty in observing the growth of hepatocytes

Method used

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  • Cell reactor and artificial liver support system comprising same
  • Cell reactor and artificial liver support system comprising same
  • Cell reactor and artificial liver support system comprising same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027]The composition of embodiment 1 cell reactor

[0028] 1. The composition of the cell reactor

[0029] Such as figure 1 As shown, the cell reactor of the present invention comprises a container 10, the bottom of the container 10 is provided with a liquid outlet 8, the top of the container is provided with a cover 3, the cover 3 is provided with a liquid inlet 2, the bottom of the container and the top of the container are sealed A layer of 100-400-mesh filter screen 7, 4 is arranged inside the cover, and the liquid outlet 8 and the inlet 2 are provided with a cover 9, 1 that is tightly matched with it, and the microcarrier with liver cells is accommodated between the two layers of filter screen inside the container. A mixture of 6 and resin 5.

[0030] The cell reactor of the present invention is loaded with a mixture of microcarriers with hepatocytes and resin, wherein the hepatocytes are animal or human primary hepatocytes, immortalized human or animal hepatic cell li...

Embodiment 2

[0032] The preparation method of embodiment 2 cell reactor

[0033] Take Cytopore 2 microcarriers (purchased from Pharmacia, USA), and pretreat before use according to the manufacturer's instructions. Cells suitable for bioartificial liver, such as immortalized human liver cell lines, are suspended in serum-free medium Keratinocyte-SFM (Invitrogen, USA), to which rEGF (final concentration 0.1ng / ml) and BPE (bovine pituitary extract )20~30μg / ml). According to the human hepatic cell line 5 × 10 7 The ratio of cells: 1g microcarrier: 500ml culture medium was mixed, cultured under shaking conditions, so that the cells were attached to the microcarrier. Change the medium every 2 days. The morphology and number of hepatocytes on the microcarriers were observed by Gimsa staining. When the cells on the microcarriers grow nearly confluent (80-100%), they are mixed with sterilized resin particles with a diameter of 800 μm (purchased from Zhuhai Livzon Medical Biomaterials Co., Ltd.)...

Embodiment 3

[0034] Embodiment 3: the regeneration method of cell reactor

[0035] When a treatment is completed, the cell reactor of the biological hepatic circulation part is removed, the bottom outlet is closed, and the serum-free cell culture medium is poured into the top inlet port, and the bottom outlet is opened after filling, so that the liquid flows from The bottom outlet flows out, repeat 3 to 5 times to fully remove the dialysate. Then, after the liquid has completely flowed out, the outlet end is closed with a cover, the cell culture medium is added to the cell reactor, the inlet end is covered with a cover, and the cover is unscrewed to allow air to pass through, and then placed in a cell incubator for cultivation; or After the dialysate is completely washed, the hepatocytes in the cell reactor are perfused with culture medium by a peristaltic pump. After regeneration and culture for 12-24 hours, a small amount of microcarriers were taken, and cell survival was observed by Gi...

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Abstract

The invention provides a cell reactor and an artificial liver support system comprising the same. The cell reactor comprises a container, a liquid outlet is formed at the bottom of the container, a sealing cover is arranged at the top of the container, a liquid inlet is formed on the sealing cover, layers of filter meshes of 100-400 meshes are respectively arranged at the bottom of the container and in the sealing cover at the top of the container, covers which are in seal fit with the liquid outlet and the inlet are formed at the liquid outlet and the inlet, and a mixture of a micro-carrier growing liver cells and resin is accommodated between the two layers of the filter meshes in the container. The reactor is connected on a dialysate circulation path of a blood purifying device for constituting the artificial liver support system, thereby leading the liver cells not to be in direct contact with blood and realizing good biological safety; furthermore, the reactor can be used for a plurality of times through regeneration treatment, thereby greatly reducing the treatment cost of a biological artificial liver.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and specifically relates to a cell reactor, a preparation method and a regeneration method thereof, and an artificial liver support system including the cell reactor. Background technique [0002] Liver failure is the end-stage manifestation of various liver diseases, and its incidence and mortality are high. Artificial liver support system is a convenient and effective treatment at present. It can provide liver function support treatment for patients, so that patients can successfully transition to Liver transplantation or restoration of damaged liver function. There are three types of artificial livers: non-biological artificial liver, biological artificial liver and hybrid artificial liver. Among them, the non-biological artificial liver includes hemodialysis, plasma exchange, activated carbon adsorption, etc. Although it can play a certain role in detoxification, it is far from the real ...

Claims

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Application Information

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IPC IPC(8): A61M1/36C12M3/00
Inventor 房青徐梅高福云张文健娄晋宁
Owner 苏州瑞徕生物科技有限公司
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