Method for producing paclitaxel by using Chinese yew cambium stem cells

A taxus and cambium technology, applied in the field of cell culture of medicinal plants, can solve the problems of low unit cell content, unstable taxus cells, etc., and achieve the effects of high taxol content, avoiding growth inhibition, and improving growth rate

Inactive Publication Date: 2019-02-12
覃家日
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the instability of yew cells during the scale-up pr

Method used

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  • Method for producing paclitaxel by using Chinese yew cambium stem cells
  • Method for producing paclitaxel by using Chinese yew cambium stem cells

Examples

Experimental program
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Effect test

Embodiment 1

[0024] A method for producing paclitaxel using yew cambium stem cells, comprising the following steps:

[0025] 1) Select the buds on the tender stems of Taxus chinensis as culture materials, wash them with detergent and tap water, rinse them under running water for 10 minutes, place them in 75% alcohol for 10 seconds, and then pass them through 0.1% liters of water. After mercury disinfection, rinse with sterile water, cut off the discolored part of the explant, and then conduct induction culture, select fresh embryogenic callus that has been subcultured for 3 times and has vigorous growth, loose texture, stable and uniform state, and inoculated into the liquid suspension. In the culture medium, shake culture, subculture once every 7 days, and establish a stable growth yew cambium stem cell culture system after subculture for 2 months;

[0026] 2) Add the yew endophytic fungus extract to the suspension culture medium, mix evenly to obtain a mixed culture solution, place the m...

Embodiment 2

[0031] A method for producing paclitaxel using yew cambium stem cells, comprising the following steps:

[0032]1) Select the buds on the tender stems of Taxus chinensis as culture materials, wash them with detergent and tap water, rinse them under running water for 10 minutes, place them in 75% alcohol for 10 seconds, and then pass them through 0.1% liters of water. After mercury disinfection, rinse with sterile water, cut off the discolored part of the explant, and then conduct induction culture, select fresh embryogenic callus that has been subcultured for 3 times and has vigorous growth, loose texture, stable and uniform state, and inoculated into the liquid suspension. In the culture medium, shake culture, subculture once every 7 days, and establish a stably growing taxus cambium stem cell culture system after 2 months of subculture; wherein, a germination bed is set in the culture dish, and the germination bed is from the top The first layer to the bottom is filter paper ...

Embodiment 3

[0038] A method for producing paclitaxel using yew cambium stem cells, comprising the following steps:

[0039] 1) Select the buds on the tender stems of Taxus chinensis as culture materials, wash them with detergent and tap water, rinse them under running water for 10 minutes, place them in 75% alcohol for 10 seconds, and then pass them through 0.1% liters of water. After mercury disinfection, rinse with sterile water, cut off the discolored part of the explant, and then conduct induction culture, select fresh embryogenic callus that has been subcultured for 3 times and has vigorous growth, loose texture, stable and uniform state, and inoculated into the liquid suspension. In the culture medium, shake culture, subculture once every 7 days, and establish a stably growing taxus cambium stem cell culture system after 2 months of subculture; wherein, a germination bed is set in the culture dish, and the germination bed is from the top The first layer to the bottom is filter paper...

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Abstract

The invention provides a method for producing paclitaxel by using Chinese yew cambium stem cells, comprising the following steps: 1)establishing a stable Chinese yew cambium stem cell culture system;2) adding a Chinese yew endophytic fungus extract solution into a suspension culture medium, inoculating Chinese yew cells, performing aerobic culture for 6-8 days under the conditions of artificial illumination, and adding methyl jasmonate and clinacanthus nutans extract to a liquid suspension medium to continue culturing; 3) collecting the Chinese yew cells, and performing crude extraction of paclitaxel; 4) purifying the crude extract; 5) adding a tetrahydrofuran-acetone mixed solution to the extract II, heating and dissolving, cold-cutting to room temperature, performing natural crystallization, re-dissolving the crystal into the tetrahydrofuran-acetone mixed solution for recrystallization for 2 to 3 times for obtaining high-purity paclitaxel. The method for producing paclitaxel by using Chinese yew cambium stem cells has the characteristics of high induction success rate and high paclitaxel content in unit cell.

Description

technical field [0001] The invention relates to the technical field of cell culture of medicinal plants. More specifically, the present invention relates to a method for producing paclitaxel using yew cambium stem cells. Background technique [0002] Taxus is a class of trees and shrubs belonging to the genus Taxus in the family Taxaceae, and there are 11 species in the world. Yew has a wide distribution range, from the subarctic regions of Eurasia and North America to the subtropical and even tropical regions of Central America and Southeast Asia. Studies have shown that paclitaxel in yew has special effects on the treatment of ovarian cancer, lung cancer, colon cancer, metastatic breast cancer, melanoma, leukemia and other cancers, making paclitaxel one of the most popular anticancer drugs at present. However, in the prior art, the source of paclitaxel is obtained from the bark of Taxus chinensis, and the content is very low, which cannot satisfy people's increasing dema...

Claims

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Application Information

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IPC IPC(8): C12N5/04C12P17/02
CPCC12N5/04C12P17/02
Inventor 覃家日
Owner 覃家日
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