Culture medium for culturing hepatocytes and preparation method thereof

A medium and hepatocyte technology, applied in biochemical equipment and methods, culture process, tissue culture, etc., can solve the problems of unfavorable daily application of hepatocyte industrial culture, low utilization rate of medium, high price, etc., and achieve stem cell Good growth, enhanced uptake and utilization, and adhesion-promoting effects

Inactive Publication Date: 2017-10-03
QINGDAO AMA CO LTD
View PDF6 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chinese patent application CN105087465A discloses a hepatocyte serum-free medium, which includes the following components: basal medium 500mL; sericin 0.05-0.5%; dexamethasone 0.1-1000nmol/mL; hepatocyte growth factor 5-20ng /mL; epidermal growth factor 10 ~ 50ng/mL; penicillin and streptomycin 100U/mL, these serum-free media are used for hepatocyte culture, but they are expen

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0023] Example 1

[0024] The invention provides a culture medium for culturing hepatocytes, comprising basal medium F12 medium and additives added in the basal medium, the ingredients of the additives include: cholesterol 6mg / L, sulfur Caprylic acid 60μg / L, chondroitin sulfate 0.3g / L, antibiotic 15μM, insulin 1.2mg / L, catalase 14mg / L, all-trans retinoic acid 4μg / L, fibronectin 16μg / L, lipid-type bovine Serum protein 300 mg / L, 2-mercaptoethanol 30 μg / L, rapamycin 6 μg / L, folic acid 5 μg / L, progesterone 1.0 mg / L, trace elements 15 μg / L, cell growth factor 30 μg / L; The growth factor is composed of epidermal growth factor and hepatocyte growth factor, and the cell growth factor is composed of epidermal growth factor and hepatocyte growth factor in a weight ratio of 1:2.

[0025] The trace elements are iron, copper, zinc, cobalt, manganese;

[0026] The additive also contains: L-glutamine, adenine;

[0027] The present invention also provides a preparation method for the medium...

Example Embodiment

[0031] Example 2

[0032]The present invention provides a culture medium for hepatocytes, comprising basal medium RMPI 1640 medium and additives added in the basal medium, the ingredients of the additives include: cholesterol 5mg / L, in terms of final concentration, Lipoic acid 42μg / L, chondroitin sulfate 0.2mg / L, antibiotic 13μM, insulin 0.9mg / L, catalase 10mg / L, all-trans retinoic acid 3μg / L, fibronectin 15μg / L, lipid type Bovine serum albumin 240 mg / L, 2-mercaptoethanol 25 μg / L, rapamycin 5 μg / L, folic acid 4 μg / L, progesterone 0.7 mg / L, trace elements 5 μg / L, cell growth factor 20 μg / L; The cell growth factor is composed of epidermal growth factor and hepatocyte growth factor, and the cell growth factor is composed of epidermal growth factor and hepatocyte growth factor in a weight ratio of 1:1.2.

[0033] The trace elements iron, copper, vitamin A, vitamin D;

[0034] The additive also contains: L-glutamine, adenine, guanine, uracil;

[0035] The present invention also ...

Example Embodiment

[0039] Example 3

[0040] The invention provides a medium for cultivating hepatocytes, comprising basal medium F12 medium and additives added in the basal medium, the ingredients of the additives include: cholesterol 8mg / L, sulfur Caprylic acid 82 μg / L, chondroitin sulfate 0.5 mg / L, antibiotic 16 μM, insulin 1.3 mg / L, catalase 17 mg / L, all-trans retinoic acid 6 μg / L, fibronectin 18 μg / L, lipid-type bovine Serum protein 360 mg / L, 2-mercaptoethanol 42 μg / L, rapamycin 8 μg / L, folic acid 7 μg / L, progesterone 1.3 mg / L, trace elements 23 μg / L, cell growth factor 35 μg / L; The growth factor is composed of epidermal growth factor and hepatocyte growth factor, and the cell growth factor is composed of epidermal growth factor and hepatocyte growth factor in a weight ratio of 1:3.6.

[0041] The trace elements are vitamins, and the vitamins are vitamin A, vitamin D, vitamin E, vitamin K, vitamin B1, vitamin B2;

[0042] The additive also contains: L-glutamine, adenine;

[0043] The pre...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention relates to the technical field of a cell culture medium and particularly relates to a culture medium for culturing hepatocytes and a preparation method thereof. The culture medium comprises a basic culture medium and additives in the basic culture medium. The additives comprise, on the basis of final concentration, 2 to 10 milligrams per liter of cholesterol, 30 to 100 micrograms per liter of lipoic acid, 0.2 to 0.5 milligrams per liter of chondroitin sulfate, 13 to 16 micromoles per liter of antibiotic, 0.9 to 1.3 milligrams per liter of insulin, 10 to 17 milligrams per liter of catalase, 2 to 7 micrograms per liter of all-transretinoic acid, 14 to 22 micrograms per liter of fibronectin, 180 to 420 milligrams per liter of fat-type bovine serum protein, 10 to 60 micrograms per liter of 2-mercaptoethanol, 3 to 12 milligrams per liter of rapamycin, 3 to 9 milligrams per liter of folic acid, 0.2 to 1.6 milligrams per liter of progesterone, 5 to 23 milligrams per liter of trace elements and 20 to 35 milligrams per liter of a cell growth factor. In the culture medium, stem cells can well grow and cell form and density are the same to those in a serum culture medium. The culture medium solves the problems of the traditional serum-containing culture medium. The culture medium has a reasonable formula. The components produce synergism, provide enough nutrients and a good environment for cell grow and proliferation and improve viability of hepatocytes.

Description

technical field [0001] The invention relates to the technical field of cell culture medium, in particular to a culture medium for culturing hepatocytes and a preparation method thereof. Background technique [0002] Using animal cell culture technology to verify the most efficient and feasible method of compound toxicology experimental testing so far. With the continuous expansion of the application field and market demand of biotechnology in the toxicological safety evaluation of compounds, improving animal cell culture and toxicology tests and extending the maintenance time to obtain reliable results have become the core of optimizing animal cell culture processes and toxicology experiments. [0003] As we all know, the economy and efficiency of animal cell culture and toxicology test are determined by the medium, and the optimization of the medium is the core link in the establishment of efficient animal cell culture and toxicology test process. Therefore, it is an impor...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12N5/071
CPCC12N5/067C12N2500/12C12N2500/30C12N2500/32C12N2500/38C12N2500/44C12N2501/10C12N2501/33C12N2501/71C12N2501/998
Inventor 翟岱青
Owner QINGDAO AMA CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap