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E.wushanense suspension cell culture system and establishment method and application thereof

A suspension cell and culture system technology, applied in the field of Wushan Epimedium suspension cell culture system, can solve the problems that wild resources cannot meet the demand and the demand for Epimedium is large

Inactive Publication Date: 2011-01-19
黎云祥
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Along with the industrialization process of modern Chinese medicine, the in-depth research of modern science on Epimedium, and the continuous discovery of new curative effects, Chinese patent medicines and ethnic medicine products with Epimedium as the main raw material continue to come out, and the market demand for Epimedium is increasing. It is getting bigger and bigger, making its wild resources gradually in danger of not being able to meet the demand
There are few reports on epimedium cell suspension culture and its flavonoid content at home and abroad

Method used

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  • E.wushanense suspension cell culture system and establishment method and application thereof
  • E.wushanense suspension cell culture system and establishment method and application thereof
  • E.wushanense suspension cell culture system and establishment method and application thereof

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Embodiment 1

[0021] Establishment of Suspension Cell Culture System of Epimedium Wushanense

[0022] The present invention adopts orthogonal test and single factor test to study the effects of inoculation amount, hormone ratio, sugar concentration and medium type on cell growth and flavonoid content to establish a suspension cell culture system of Epimedium wushanense.

[0023] 1. Materials and methods

[0024] 1.1 Material culture The callus induced from the young leaves of Epimedium herba is selected as the inoculation material with uniform growth, loose texture and light yellow callus after repeated screening after subculture. After 16 days of callus inoculation and subculture, the inoculation was carried out in the culture medium for shaking culture (120rpm), and the light was 12h·d -1 . The culture medium adopts the same hormone and sucrose concentration ratio as the culture medium of subcultured callus, LS medium +2, 4-D2.0mg·L -1 +0.5mg·L -1 6-BA+Sugar30g·L -1 , just without th...

Embodiment 2

[0063] Growth Experiment of Cells in Suspension Culture System of Epimedium Wushanense

[0064] Instruments: Leici Phs-25 digital display pH meter, Shanghai Precision Scientific Instrument Co., Ltd.; LDZX-40BI vertical automatic pressure steam sterilizer, Shanghai Shen’an Medical Instrument Factory; HZQ-QX full-temperature oscillator, Harbin Donglian Electronic Technology Development Co., Ltd.; GBC 916 UV-Vis Spectrophotometer (produced by GBC Scientific Instruments Co., Ltd., Australia)

[0065] Reagent: Icariin reference substance (purchased from China Institute for the Control of Pharmaceutical and Biological Products).

[0066] Culture system: The culture system is based on the callus obtained from the young leaves of Epimedium wushanense (Guizhou medicinal plant-Epimedium taxonomy expert He Shunzhi, identified by Teacher Qin Ma Yonghong of China West Normal University) in the laboratory, and after several generations of cultivation The obtained Epimedium wushanense homog...

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Abstract

The invention discloses an E.wushanense suspension cell culture system and an establishment method and application thereof. The system is characterized by adopting the orthogonal test and the single factor test to study the impacts of the inoculum size, the hormone combination, the sugar concentration and the culture media variety on the cell growth and the flavonoid content. The ideal E.wushanense suspension cell culture system can be easily established by utilizing the crisp embryonic calli. The E.wushanense callus cell undergoes suspension culture in the B5 basic culture medium, 1.0mg.1-12, 4-D and 0.2mg.1-1BA are added, the sugar concentration is 30-50g.1-1 and the inoculum size is 2g of fresh weight per 30ml, thus the established suspension cell culture system can obtain high-yield flavonoids.

Description

technical field [0001] The invention relates to a suspension cell culture technology, in particular to a Wushan epimedium suspension cell culture system and its establishment and application method. Background technique [0002] Wushan Epimedium (E. wushanense) is a perennial herbaceous plant derived from the genus Epimedium of the family Berberdiaceae. In Li Shizhen's "Compendium of Materia Medica", it is said that it has the effect of "benefiting essence, strengthening muscles and bones, strengthening waist and knees, and strengthening heart". Modern pharmacological experimental studies have shown that Epimedium can increase blood flow in cardiovascular and cerebrovascular, promote hematopoietic function, immune function and bone metabolism, and has anti-aging, anti-tumor, kidney-yang, strong bones, and rheumatism, etc. It has a wide range of clinical applications. National Pharmacopoeia Committee. Pharmacopoeia of the People's Republic of China (Part One) [S]. Beijing: C...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/04
Inventor 黎云祥韩素菊权秋梅
Owner 黎云祥