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Serum free hepatocyte medium

A medium and hepatocyte technology, applied in the field of serum-free hepatocyte medium, can solve the problems of high price, secret ingredient formula, and inability to culture without serum, and achieve the effect of good growth.

Active Publication Date: 2014-09-10
WEST CHINA HOSPITAL SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the HepatoZYME-SFM serum-free medium of Invitrogen Company of the United States has been listed, but Masashi Kitazawa et al. (2007) Angiopoietin-related growth factor suppresses gluconeogenesis through the akt / forkhead box calss O 1-dependent pathway in hepatocytes reported that using HepatoZYME -When cultivating primary rat hepatocytes in SFM serum-free medium, 10% fetal bovine serum still needs to be added, which cannot truly achieve serum-free culture
Structural polarity and functional bile canaliculi in rat hepatocyte spheroids. Experimental cell research 274: 56-67, 2002; Sustaining a bioatificial liver under hypothermic conditions. Tissue engineering 11: 427-437, 2005.; stellate cells. Cytotechnology 45: 125-140, 2004.; Dexamethasone effects on rat hepatocyte spheroid formation and function. Tissue engineering 11: 415-426, 2005.; etc. have disclosed three serum-free media, but these three serum-free media contain animal-derived components and cannot be used for hepatocyte suspension culture, which is difficult to meet the needs of scientific research and clinical application
Other commercial hepatocyte serum-free medium has problems such as the secret formula of the medium, and the price is expensive, which discourages the majority of hepatocyte transplantation and BAL researchers and users, and urgently needs to find a new hepatocyte serum-free culture medium. base

Method used

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Embodiment 1

[0037] Embodiment 1 Preparation of culture medium of the present invention

[0038] 1. Preparation method

[0039] (1) Take the following ingredients:

[0040]

[0041]

[0042] (2) Mix the above medium components, dissolve in deionized water, stir at room temperature for 10 minutes to fully dissolve, dilute to 1L, and sterilize.

[0043] When using the medium, use deionized water to adjust the sodium ion concentration of the medium to 1.3g / L (the allowable error range is 1.28g / L to 1.32g / L), adjust the pH to 7.2, and filter to sterilize with a 0.2 micron filter membrane , preheated to 37°C.

Embodiment 2

[0044] Embodiment 2 Preparation of culture medium of the present invention

[0045] 1. Preparation method

[0046] (1) Take the following ingredients:

[0047]

[0048] (2) Mix the above medium components, dissolve in deionized water, stir at room temperature for 10 minutes to fully dissolve, dilute to 1L, and sterilize.

[0049] When using the medium, use deionized water to adjust the sodium ion concentration of the medium to 1.3g / L (the allowable error range is 1.28g / L to 1.32g / L), adjust the pH to 7.2, and filter to sterilize with a 0.2 micron filter membrane , preheated to 37°C.

Embodiment 3

[0050] Embodiment 3 Preparation of culture medium of the present invention

[0051] 1. Preparation method

[0052] (1) Take the following ingredients:

[0053]

[0054] (2) Mix the above medium components, dissolve in deionized water, stir at room temperature for 10 minutes to fully dissolve, dilute to 1L, and sterilize.

[0055] When using the medium, use deionized water to adjust the sodium ion concentration of the medium to 1.3g / L (the allowable error range is 1.28g / L to 1.32g / L), adjust the pH to 7.2, and filter to sterilize with a 0.2 micron filter membrane , preheated to 37°C.

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Abstract

The invention provides a novel serum free medium and a preparation method and application of the medium. The serum free medium overcomes defects of the traditional serum media, and is obviously superior to serum free media reported in the conventional documents, improves the possibility of supporting systematical treatment clinical application through cell transplantation, tissue engineering liver and bioartificial liver, and has high practical value.

Description

technical field [0001] The invention relates to a hepatocyte culture medium, in particular to a serum-free hepatocyte culture medium. Background technique [0002] Liver parenchymal cells, namely hepatocytes, are the main executors of liver function, and their total weight accounts for about 80% of liver weight. Hepatocytes absorb and process nutrients and other substances in the blood, such as glucosylating bilirubin and finally forming bile to be secreted into the intestinal tract. At the same time, hepatocytes also synthesize albumin, complement proteins, glycogen (glucose storage) and coagulation In addition, liver cells also convert ammonia in the blood into urea to complete detoxification. The liver also plays an important role in the regulation of carbohydrates, lipids and amino acids. The specific functions of the liver are completed by various liver-specific enzymes synthesized by liver cells, which synthesize and store the nutrients absorbed by the gastrointestina...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071
Inventor 包骥石毓君步宏
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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