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A method for separating and culturing ginseng stem cells using a biological reaction device

A biological reaction device and a separation and culture technology, which are applied in the field of ginseng stem cell separation and culture, can solve the problems that the content of effective components of stem cells needs to be improved, the ability to divide is limited, and the ability to resist stress is not strong, so as to shorten the induction time, improve the growth, and achieve uniform ventilation. and sufficient effect

Active Publication Date: 2022-07-08
SHANDONG ANRAN NANOMETRE IND DEV CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this kind of cell essence comes from differentiated somatic cells, so the ability to divide is limited and the ability to resist stress is not strong
In industrial production, cell line degeneration is prone to occur, and the ability to divide is weak
In addition, the culture period is long, and the content of active ingredients in the obtained stem cells also needs to be improved.

Method used

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  • A method for separating and culturing ginseng stem cells using a biological reaction device
  • A method for separating and culturing ginseng stem cells using a biological reaction device
  • A method for separating and culturing ginseng stem cells using a biological reaction device

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0102] In the present embodiment, the adventitious root of ginseng is prepared by one-step method and specific liquid medium culture, which specifically includes the following steps:

[0103] (1) Induction of adventitious roots

[0104] The 20-year-old wild ginseng, removed the reed head, the stern part, the main root after cleaning, disinfection and sterilization, was cut into thin slices with a width of 0.6cm, a length of 0.7cm and a thickness of 0.3mm, and inoculated into the induction medium at 22±1°C. Under the condition of dark culture for 4 to 5 weeks, the adventitious roots of ginseng were induced; wherein, the induction medium included 4mg / L naphthalene acetic acid, 0.6mg / L gibberellin, 0.4mg / L kinetin, 0.1g / L citric acid, 0.05g / L L ascorbic acid, 30 g / L sucrose, 3 g / L phytogel, 1.55 g / L B5 medium and 1.21 g / L WPM medium, pH 5.8.

[0105] (2) Subculture of adventitious roots

[0106] Inoculate the adventitious root of the wild ginseng obtained in step (1) into the sam...

Embodiment 2

[0111] In the present embodiment, the adventitious root of ginseng is prepared by a one-step method and a specific liquid medium culture, which specifically includes the following steps:

[0112] (1) Induction of adventitious roots

[0113] After washing, sterilizing and sterilizing the head of the 6-year-old garden ginseng, cut into slices with a width of 0.5cm, a length of 0.6cm and a thickness of 0.3mm, inoculated into the induction medium, and cultivated in the dark at 22±1°C for 4-5 days. Adventitious roots were induced every week; wherein, the induction medium included 6mg / L naphthalene acetic acid, 0.2mg / L gibberellin, 0.4mg / L kinetin, 1.2g / L citric acid, 0.1g / L ascorbic acid, 20g / L sucrose, 5g / L phytogel, 4g / L B5 medium and 1.8g / LWPM medium, pH 5.6.

[0114] (2) Subculture of adventitious roots

[0115] The adventitious roots obtained in step (1) are inoculated into the same induction medium as in step (1), and cultured in the dark for 4-5 weeks under the same condit...

Embodiment 3

[0120] In the present embodiment, the adventitious root of ginseng is prepared by a one-step method and a specific liquid medium culture, which specifically includes the following steps:

[0121] (1) Induction of adventitious roots

[0122] After washing, sterilizing and sterilizing the stem of 10-year-old ginseng under the forest, cut into slices with a width of 0.7cm, a length of 0.7cm and a thickness of 0.5mm, inoculated into the induction medium, and cultivated in the dark at 22±1℃ for 4~ Induce adventitious roots in 5 weeks; wherein, the induction medium includes 5mg / L naphthalene acetic acid, 1mg / L gibberellin, 0.1mg / L kinetin, 0.75g / L citric acid, 0.03g / L ascorbic acid, 40g / L sucrose, 4g / L phytogel, 2g / L B5 medium and 1g / L WPM medium, pH 6.0.

[0123] (2) Subculture of adventitious roots

[0124] The adventitious roots obtained in step (1) are inoculated into the same induction medium as in step (1), and cultured in the dark for 4-5 weeks under the same conditions;

...

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Abstract

The invention discloses a method for separating and culturing ginseng stem cells using a biological reaction device. stem cell area; (3) inoculate the stem cells obtained by separation in step (2) into a stem cell induction medium for dark culture to obtain a stem cell mass, and then transfer the stem cell mass into a subculture medium for dark culture; (4) put the Step (3) inoculating the subcultured ginseng stem cells into the stem cell liquid culture medium shake flask, and culturing in the dark under the condition of 100-140rpm; (5) inoculating the ginseng stem cells cultured in the shake flask in step (4) into a biological reaction device medium, cultured in the dark to obtain ginseng stem cells. By adopting the liquid medium and the culturing method of the present invention, adventitious roots can be directly induced by various parts of mature ginseng, the induction steps are simplified, the induction time is shortened, and the content of ginsenosides in the adventitious roots is increased, and ginseng stem cells with good indicators are finally obtained.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, in particular to a method for separating and culturing ginseng stem cells using a biological reaction device. Background technique [0002] Ginseng (Panax ginseng C.A.Mey.) is an Araliaceae ginseng plant, distributed in China, Japan, and Korea. Its rhizomes are valuable Chinese medicinal materials, known as "the king of herbs". Ginseng is sweet, slightly bitter, slightly warm, and has the functions of invigorating vitality, strengthening the pulse, invigorating the spleen and benefiting the lung, promoting body fluid and nourishing blood, soothing the mind and nourishing wisdom, etc. Wound thirst, palpitations, insomnia, etc. Ginsenoside is the main active ingredient of ginseng, which has the functions of anti-fatigue, anti-aging, regulating the central nervous system, improving immunity, improving cardiovascular and cerebrovascular insufficiency, and inhibiting tumor cell productio...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/04C12M1/04C12M1/12C12M1/00A01H4/00
CPCC12N5/04C12M29/06C12M29/20C12M23/54C12M23/22C12M29/04A01H4/001A01H4/006
Inventor 张明臣刘冰闫培生高秀君
Owner SHANDONG ANRAN NANOMETRE IND DEV CO LTD
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