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Preparation method of ginseng stem cells

A technology of stem cells and ginseng, applied in the field of stem cells, can solve the problems of difficult to control the dosage of auxin, difficult operation, low success rate, etc., and achieve the effects of strong resistance to stress, fast growth, and reduced production costs

Active Publication Date: 2018-12-18
厦门鹭港兆康生物科技有限公司
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The purpose of the present invention is to provide a preparation method of ginseng stem cells, so as to solve the problem that it is difficult to control the dosage of auxin in the separation technology of ginseng stem cells in the prior art, which leads to difficult operation and low success rate.

Method used

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  • Preparation method of ginseng stem cells
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  • Preparation method of ginseng stem cells

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Effect test

Embodiment 1

[0027] The invention discloses a preparation method of ginseng stem cells, comprising the following steps:

[0028] Step 1. Sampling: Take fresh ginseng rhizomes, sterilize them with a 70% ethanol solution on an ultra-clean workbench for 30 seconds, then sterilize them with a 0.1% mercuric chloride solution for 10 minutes, and then Wash 3 times with sterile water; cut the ginseng rhizome into slices with a thickness of 3 mm; on the rhizome slices, the ginseng cambium presents a circle of dark yellow, such as figure 1 As shown, carefully cut along the dark yellow circle with a scalpel under a stereomicroscope, cut off the dark yellow circle, and cut it into sections with a length of 4 mm and a width of 2 mm;

[0029] Step 2. Preparation of stem cell protoplasts: Soak the above-mentioned sections in the enzymolysis solution containing the following mass fractions: 0.8% cellulase, 0.8% pectinase, 12% mannitol and 86.4% distilled water, and incubate at room temperature After 20 h...

Embodiment 2

[0036]The effect of different enzyme concentrations on the separation of protoplasts was examined, and the results are shown in Table 1. When 0.8% cellulase, 0.8% pectinase and 12% mannitol, the yield of protoplasts reached 9.8×10 3 / g fresh weight, the survival rate reached 95.3%, significantly better than other groups.

[0037] Table 1: Effect of Enzyme Concentration on Protoplast Isolation

[0038] cellulase

[0039] In a word, the present invention not only slices the ginseng root, but also microcuts the cells at the cambium into very thin slices, and then dissociates the stem cells at the cambium and the protoplasts of the surrounding body cells by enzymatic hydrolysis , and then regenerate the cell wall and obtain single-cell clones through nursing culture, and then identify stem cell lines from these single-cell clones. Although this method has many steps, it is easy to implement and has a very high probability of success.

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Abstract

The invention discloses a preparation method of ginseng stem cells, wherein the preparation method comprises: sampling, preparing stem cell protoplasts, carrying out in vitro culture, carrying out nursing culture, carrying out single cell cloning culture, and carrying out subculture. According to the present invention, with the method, the ginseng stem cells can be effectively separated and cultured, and the a new feasible way can be provided for the separation of other tuber and rhizome plant stem cells; and the cultured ginseng stem cells have characteristics of infinite division ability, fast growth and strong reverse resistance, provides the basis for super-large-volume liquid suspension culture, and has the significantly-reduced production cost.

Description

technical field [0001] The invention relates to the technical field of stem cells, in particular to a preparation method of ginseng stem cells, and can also be used for the separation method of stem cells of other tuberous plants. Background technique [0002] Ginseng is a precious Chinese medicinal material in my country, which has a long history of use and is recognized by the public. The medicinal ingredients extracted from the medicinal plant ginseng have important application value and prospects for the treatment of various diseases and nutritional health care. Due to the long growth cycle of ginseng and the occupation of cultivated land; and the production process is easily affected by environmental pollution, diseases and insect pests, the traditional ginseng planting can no longer meet the market demand. [0003] The relevant medicinal ingredients are produced by in vitro suspension culture of ginseng cells, and this method can obtain a large number of ginseng cells...

Claims

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Application Information

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IPC IPC(8): C12N5/04
CPCC12N5/04C12N2500/05C12N2500/30C12N2500/40C12N2509/00
Inventor 黄涛
Owner 厦门鹭港兆康生物科技有限公司
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