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E.wushanense suspended cell culture system, and establishing 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 of large demand for Epimedium and unsatisfactory wild resources

Inactive Publication Date: 2011-11-23
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 suspended cell culture system, and establishing method and application thereof
  • E.wushanense suspended cell culture system, and establishing method and application thereof
  • E.wushanense suspended cell culture system, and establishing method and application thereof

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Embodiment 1

[0026] Establishment of Suspension Cell Culture System of Epimedium Wushanense

[0027] 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.

[0028] 1. Materials and methods

[0029] 1.1 Material culture The callus induced from the young leaves of Epimedium wushanense was selected as the inoculation material after subculture and repeated screening. The callus with uniform growth, loose texture and good dispersion was light yellow.

[0030] 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...

Embodiment 2

[0071] Growth Experiment of Cells in Suspension Culture System of Epimedium Wushanense

[0072] 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)

[0073] Reagent: Wushan icariin reference substance (purchased from China Institute for the Control of Pharmaceutical and Biological Products).

[0074] 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 wushanens...

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Abstract

The invention discloses an E.wushanense suspended cell culture system, and an establishing method and application thereof. According to the invention, orthogonal tests and single factor tests are employed, and research on influence of inoculation amount, hormone ratio, sugar concentration and medium varieties on cell growth and flavonoid content is carried out. An ideal E.wushanense suspended cell culture system can be established easily by using crisp embryonic callus. The suspended cell culture system, established by suspension culture of E.wushanense callus cells in B5 basal medium in which 1.0 mg / L of 2,4-D and 0.2 mg / L of BA are added, with cane sugar concentration being 30 to 50 g / L and inoculation amount being 2 g by fresh weight per mL, enables high-yield flavones to be obtained.

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): C12N5/04C12N5/02
Inventor 黎云祥韩素菊权秋梅
Owner 黎云祥