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A kind of cultivation method and application of dendrobium candidum stem cells

A technology of Dendrobium officinale and a culture method, which is applied to plant cells, preparations for skin care, medical preparations containing active ingredients, etc. Significant antioxidant and anti-aging activity, obvious differentiation, rapid proliferation effect

Active Publication Date: 2018-10-02
北京添易生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, due to the inability of plant cell culture technology to maintain stable, rapid cell proliferation and high metabolites during the culture period, the application of Dendrobium officinale anti-aging plant stem cells is limited.
In order to solve this problem, Chinese patent application CN201410486185.1 discloses a stem cell of Dendrobium officinale and a method for its isolation and culture. The method is to start culturing from the cell line of the quiescent center of the root tip of Dendrobium officinale to obtain stable genetic plant stem cells for cultivation. The culture method solves the problem of easy browning of Dendrobium candidum in vitro materials. Because the cells of its stem cell line have not yet differentiated, it has the advantage of strong cell activity; however, the stem cells in the quiescent center have a longer cell cycle, and cell culture requires a longer period of time. Time, can not maintain stable, faster cell proliferation, can not meet the industrial production of plant cell culture technology

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  • A kind of cultivation method and application of dendrobium candidum stem cells
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Experimental program
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Embodiment 1

[0017] The present invention will be further described in detail below. It should be pointed out that the following description is only an illustration of the technical solutions claimed in the present invention, and is not any limitation to these technical solutions. The protection scope of the present invention shall be determined by the contents described in the appended claims. Embodiment 1 The cultivation of Dendrobium officinale stem cells

[0018] 1) Take the root tip of Dendrobium officinale with the root cap removed, wash it with water for 3 hours, then disinfect it with 75% alcohol for 15 seconds, rinse it with sterile water for 4 times, then disinfect it with 1% benomyl solution for 10 minutes, and rinse it with sterile water for 4 minutes. time, spare;

[0019] 2) Cut the root tip of Dendrobium candidum obtained in step 1 into 5mm small pieces, inoculate it on the screening medium, cultivate it at 25-29° C., and culture it in the dark for 5-10 days to obtain the ...

Embodiment 2

[0022] Example 2 Investigation on the Yield of Dendrobium officinale Stem Cells under Different Medium Conditions

[0023] According to the method of embodiment 1, get the root tip of Dendrobium candidum of equal amount and carry out stem cell culture, difference is only to adopt different screening medium and liquid medium, specifically as follows:

[0024]

[0025] The obtained stem cells of Dendrobium officinale were cryogenically frozen for 3 hours, then ultrasonically crushed and freeze-dried to obtain dry powder of stem cells of Dendrobium officinale. The weight of the dry powder was weighed each time, and each group was tested 5 times in parallel, and the average value was taken. The specific results are as follows:

[0026]

Embodiment 3

[0027] Example 3 Investigation of Antioxidative Activity of Dendrobium officinale Stem Cells under Different Medium Conditions

[0028] Take human dermal fibroblasts (HDF) for culture. When the human dermal fibroblasts grow to 80% confluence, the control group wraps the cell culture flask with tin foil, and irradiates the HDF cells with UVA ultraviolet rays for 2 hours a day. , irradiated for 4 days. Observing the morphology of HDF cells under a microscope, it was found that HDF cells began to shrink, and individual cells were floating in the culture medium; on the fifth day, the administration group was added with different groups of Dendrobium officinale stem cell dry powder with a final concentration of 10 μg / ml, and the model The group was given the same amount of culture medium, continued to culture for 48 hours, and collected HDF cells; the lysate was lysed, centrifuged at 4°C for 10 minutes, and the supernatant was taken to measure SOD activity with a SOD detection kit....

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Abstract

The invention belongs to the technical field of biology, and particularly relates to a cultivation method and application of dendrobium officinale stem cells. The cultivation method has the advantages of being remarkable in differentiation, rapid in multiplication and short in cycle. The prepared dendrobium officinale stem cells have remarkable oxygen resistance and aging resistance activity and can be widely applied to the fields of medicine, cosmetics and health-care products.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for culturing stem cells of Dendrobium officinale and an application thereof. Background technique [0002] Dendrobium officinale Kimura et Migo is a perennial epiphytic herbaceous plant belonging to the genus Dendrobium in the family Orchidaceae, also known as black knot grass. Dendrobium officinale is dried and processed into a spiral body, called "Tiepi Fengdou". Dendrobium officinale is distributed in Southwest my country and Jiangnan provinces, and mostly epiphytes in damp tree trunks or rock crevices. Due to the extremely low reproduction rate and survival rate of Dendrobium officinale under natural conditions and the long growth cycle, wild Dendrobium officinale is on the verge of extinction. [0003] Plant stem cells, also known as meristems, are the source of plant growth and development and the center of signal regulation. Since plant stem cells are ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/04A61K36/8984A61K8/97A61P39/06A61Q19/08
Inventor 韩晓菲林倩如苏永军
Owner 北京添易生物科技有限公司
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