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Ginseng stem cell isolated culture method using biological reaction device

A biological reaction device and a technology for separation and culture, 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 resist stress is not strong, and the ability to divide is limited, so as to achieve shortened induction time, uniform and sufficient ventilation, The effect of increasing the content of active ingredients

Active Publication Date: 2021-07-23
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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  • Ginseng stem cell isolated culture method using biological reaction device
  • Ginseng stem cell isolated culture method using biological reaction device
  • Ginseng stem cell isolated culture method using biological reaction device

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0103] In this example, the adventitious root of ginseng is prepared by one-step method and specific liquid medium culture, which specifically includes the following steps:

[0104] (1) Induction of adventitious roots

[0105] After 20-year-old wild ginseng, remove the reed head, stalk, and main root after cleaning, disinfection and sterilization, cut into thin slices with a width of 0.6cm, a length of 0.7cm and a thickness of 0.3mm, and inoculate them into induction medium, at 22±1°C Adventitious roots of wild ginseng were induced by dark culture for 4 to 5 weeks under conditions; wherein, the induction medium included 4mg / L naphthaleneacetic acid, 0.6mg / L gibberellin, 0.4mg / L kinetin, 0.1g / L citric acid, 0.05g / L L ascorbic acid, 30g / L sucrose, 3g / L Phytogel, 1.55g / L B5 medium and 1.21g / L WPM medium, pH value is 5.8.

[0106] (2) Subculture of adventitious roots

[0107] Inoculate the adventitious roots of wild ginseng obtained in step (1) into the same induction medium as ...

Embodiment 2

[0112] In this example, the adventitious root of ginseng is prepared by one-step method and specific liquid medium culture, which specifically includes the following steps:

[0113] (1) Induction of adventitious roots

[0114] After cleaning, disinfecting and degerming the reed heads of 6-year-old garden ginseng, cut them into thin slices with a width of 0.5 cm, a length of 0.6 cm and a thickness of 0.3 mm, inoculate them into induction medium, and culture them in dark at 22±1°C for 4-5 days. Zhou induced adventitious roots; wherein, the induction medium included 6mg / L naphthaleneacetic acid, 0.2mg / L gibberellin, 0.4mg / L kinetin, 1.2g / L citric acid, 0.1g / L ascorbic acid, 20g / L sucrose, 5g / L plant gel, 4g / L B5 medium and 1.8g / L WPM medium, pH value is 5.6.

[0115] (2) Subculture of adventitious roots

[0116] Inoculate the adventitious roots obtained in step (1) into the same induction medium as in step (1), and culture in dark under the same conditions for 4 to 5 weeks;

...

Embodiment 3

[0121] In this example, the adventitious root of ginseng is prepared by one-step method and specific liquid medium culture, which specifically includes the following steps:

[0122] (1) Induction of adventitious roots

[0123] After cleaning, disinfecting and sterilizing the horns of 10-year-old understory ginseng, cut them into thin slices with a width of 0.7 cm, a length of 0.7 cm, and a thickness of 0.5 mm, inoculate them into induction medium, and culture them in dark at 22±1°C for 4~ Adventitious roots were induced in 5 weeks; wherein, the induction medium included 5mg / L naphthaleneacetic 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.

[0124] (2) Subculture of adventitious roots

[0125] Inoculate the adventitious roots obtained in step (1) into the same induction medium as in step (1), and culture in dark under the same conditions for 4 to 5 weeks;

[0126...

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Abstract

The invention discloses a ginseng stem cell isolated culture method using a biological reaction device. The isolated culture method comprises the following steps: (1) preparing ginseng adventitious roots by adopting a one-step method; (2) taking root tip parts of the ginseng adventitious roots prepared in the step (1), and performing dissection and separation to obtain a stem cell region; (3) inoculating stem cells obtained by the separation in the step (2) into a stem cell induction medium for dark culture to obtain a stem cell mass, and then transferring the stem cell mass into a subculture medium for dark culture; (4) inoculating ginseng stem cells subcultured in the step (3) into a stem cell fluid medium-contained shake flask, and performing dark culture under the condition of 100-140 rpm; and (5) inoculating the ginseng stem cells after being subjected to shake flask culture in the step (4) into the biological reaction device, and performing dark culture to obtain ginseng stem cells. By adopting a fluid medium and the culture method disclosed by the invention, the adventitious roots can be directly generated through induction of all parts of mature ginseng, the content of ginsenoside in the adventitious roots is increased while the induction step is simplified and the induction time is shortened, and finally, the ginseng stem cells with good indexes are obtained.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, and in particular relates 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 a plant of the genus Panax of Araliaceae, distributed in China, Japan, and Korea. Its rhizome is a valuable Chinese medicinal material, known as the "King of Herbs". Ginseng is sweet, slightly bitter, and slightly warm. It has the functions of invigorating vital energy, restoring pulse and strengthening the spleen, nourishing the spleen and lung, promoting body fluid and nourishing blood, calming the nerves and improving intelligence, etc. Wound thirst, palpitations, insomnia, etc. Ginsenoside is the main active ingredient of ginseng, which has the effects of anti-fatigue, anti-aging, regulating the central nervous system, improving immunity, improving cardiovascular and cerebrovascular insufficien...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/04C12M3/00C12M1/12C12M1/04
CPCC12N5/04C12N5/0025C12M23/22C12M23/38C12M23/48C12M29/04C12M29/20
Inventor 闫培生张明臣高秀君刘冰
Owner SHANDONG ANRAN NANOMETRE IND DEV CO LTD
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