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Amplified culture method of siraitia grosvenorii suspension cells

A technology of amplifying and suspending cells, applied in cell culture medium, plant cells, tissue culture, etc., can solve the problems of staying in shake flasks, unable to carry out large-scale culture, low cell yield, etc., and achieve the effect of promoting synthesis

Active Publication Date: 2020-01-14
EAST CHINA UNIV OF SCI & TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It overcomes the problems that the existing Luo Han Guo suspension culture stays at the level of shake flasks, the cell yield is low, and it is impossible to carry out large-scale culture.

Method used

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  • Amplified culture method of siraitia grosvenorii suspension cells
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  • Amplified culture method of siraitia grosvenorii suspension cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Step 1: Establishment of Luo Han Guo Cell Liquid Suspension Culture System

[0031] The mature seed embryos of Luo Han Guo were selected and disinfected with 70% alcohol and sodium hypochlorite solution containing 2.5% available chlorine under aseptic conditions. After disinfection, they were rinsed with distilled water for 5 times to clean the disinfectant. After peeling off the outer hard seed shell of the seed embryo, cut out the wound and add it to solid B5 medium containing hormone to induce embryogenic loose callus. The solid medium is composed of B5 medium with 0.5mg / L6-BA, 0.5mg / LNAA, 10g / L sucrose, 50mg / L inositol, and 3g / L agar. After 20 days of induction, select the ones with loose embryogenicity, vigorous growth, and state. The more uniform callus was subcultured.

[0032] The embryogenic loose callus that was subcultured 5 times and in a relatively stable state was inserted into the liquid medium, and the inoculation amount was 25 g / L of wet weight. Durin...

Embodiment 2

[0036] Step 1: Establishment of Luo Han Guo Cell Liquid Suspension Culture System

[0037]The mature seed embryos of Luo Han Guo were selected and disinfected with 70% alcohol and sodium hypochlorite solution containing 2.5% available chlorine under aseptic conditions. After disinfection, they were rinsed with distilled water for 6 times to clean the disinfectant. After peeling off the outer hard seed shell of the seed embryo, cut out the wound and add it to solid B5 medium containing hormone to induce embryogenic loose callus. The solid medium is composed of B5 medium with 1.5mg / L6~BA, 1.5mg / LNAA, 50g / L sucrose, 150mg / L inositol, 6g / L agar, and after induction for 30 days, select the ones with loose embryogenicity, vigorous growth and state. The more uniform callus was subcultured.

[0038] The embryogenic loose callus that has been subcultured for 15 times and in a relatively stable state is inserted into the liquid medium, and the inoculation amount is 100 g / L of wet weigh...

Embodiment 3

[0042] Step 1: Establishment of Luo Han Guo Cell Liquid Suspension Culture System

[0043] The mature seed embryos of Luo Han Guo were selected and disinfected with 70% alcohol and sodium hypochlorite solution containing 2.5% available chlorine under aseptic conditions. After disinfection, they were rinsed with distilled water for 5-6 times to clean the disinfectant. After peeling off the outer hard seed shell of the seed embryo, cut out the wound and add it to solid B5 medium containing hormone to induce embryogenic loose callus. The solid medium composition is B5 medium supplemented with 1.0mg / L6-BA, 1.0mg / LNAA, 30g / L sucrose, 100mg / L inositol, and 4.6g / L agar.

[0044] The callus with loose embryogenicity, vigorous growth and relatively uniform state was induced for subculture on day 25.

[0045] The embryogenic loose callus that has been subcultured for 10 times and in a relatively stable state is inserted into the liquid medium, and the inoculation amount is 50 g / L of we...

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Abstract

The invention provides an amplified culture method of siraitia grosvenorii suspension cells. According to the method, two or more of phenylalanine, methyl jasmonate and betaine are added in the amplified culture process of the siraitia grosvenorii suspension cells. According to the amplified culture method disclosed by the invention, the shortcoming that conventional siraitia grosvenorii suspension culture rests on a shake flask level, the cell yield is low and large-scale culture cannot be performed is overcome, the most optimal optimization culture medium for promoting product synthesis andan addition technology are obtained through screening, the synthesis rate of mogroside V is substantially increased, and a definite technical method and base are provided for larger-scale siraitia grosvenorii cell suspension culture.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for amplified culture of monk fruit suspension cells. Background technique [0002] Luo Han Guo (Siraitia grosvenorii) is derived from the genus Momordica of Cucurbitaceae, and is a precious medicinal and edible plant unique to Guilin, Guangxi. Mogroside is the main active ingredient in Luo Han Guo, which has the functions of clearing away heat and nourishing the lungs, relieving the throat and opening the tone, moistening the intestines and defecating. Recent studies have shown that Luo Han Guo also has antioxidant, anti-diabetic and anti-cancer effects. Luo Han Guo is deeply loved by people for its novel taste, pure taste and unique health functions. It is the best sweetener and health product for patients with obesity, high blood pressure and diabetes, and has a wide range of application values. The existing methods for obtaining mogroside V are mainly through extractio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/04C12N5/02
CPCC12N5/04C12N5/0025
Inventor 郭美锦宋云飞李佳瑞王泽建庄英萍杨文国蒋治舟
Owner EAST CHINA UNIV OF SCI & TECH
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