CN105400731A - Taxus chinensis cell culture medium promoting paclitaxel excretion - Google Patents
Taxus chinensis cell culture medium promoting paclitaxel excretion Download PDFInfo
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- CN105400731A CN105400731A CN201510964059.7A CN201510964059A CN105400731A CN 105400731 A CN105400731 A CN 105400731A CN 201510964059 A CN201510964059 A CN 201510964059A CN 105400731 A CN105400731 A CN 105400731A
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Abstract
The invention provides a taxus chinensis cell culture medium promoting paclitaxel excretion. According to the culture medium, a traditional autotrophic culture medium is adopted for providing a nitrogen source and a phosphorus source, a certain quantity of calcium ions are added, and on this basis, it is unexpectedly found that the yield of paclitaxel can be increased by adding specific types of traditional Chinese medicinal materials with specific ingredients. Based on the helpful finding, roots of wisteflower wisteria, sesamum indicum seeds, tinospora sinensis(Lour.)Merr. common bluebeard herb, gynura japonica, stephania sinica diels, pogonatherum crinitum, saffron crocus, flos imperatae and other traditional Chinese medicinal materials originally used for treating human diseases are preferably selected through an experimental means, an outstanding culture effect of the obtained culture medium is unexpectedly achieved, the paclitaxel yield of taxus chinensis cells is remarkably increased, and meanwhile the paclitaxel production cycle is shorter. Excellent technical effects are achieved by the innovative technological means, cost is low, and realization is easy, so that the culture medium has good application prospects.
Description
Technical field
The present invention relates to technical field of cell culture, be specifically related to a kind of Taxus chinensis cell cultures base promoting taxol to secrete.
Background technology
Ramulus et folium taxi cuspidatae, also known as Japanese yew, also claims red Bai Song, is China national one-level rare and protected trees, is also the endangered natural rare plants for anticancer of generally acknowledging in the world.
In the Taxus that taxol and analogue thereof exist only in gymnosperm taxaceae up to now and Austrotaxus, the Chinese yew genus plants whole world has 11 kinds, be distributed widely in the frigid zone of Eurasia and North America, temperate zone and subtropical zone, but the scattered and poor growth of distributing.Austrotaxus is only distributed in the Southern Hemisphere, rare numbers, and taxol content in Chinese yew is general very low, even if also less than 0.1% in the highest yewtree bark of current generally acknowledged content, but it is in great demand.
In order to solve the sharp contradiction between taxol supply and demand, correlative study person has carried out large quantity research in searching and expansion medicine source of Taxol approach for many years.Obtain successfully though the chemistry of taxol is complete synthesis, because the reasons such as route is long, yield is low cannot realize industrialization at present.In recent years, utilize yew cell isolated culture to obtain increasing concern to prepare taxol, because the training method of in vitro cell can obtain more various control, therefore its production efficiency is relatively high.But, be not easy by the cell cultures prepared for the purpose of taxol, first need to screen the strong cell strain of production performance, also to mate suitable culture condition simultaneously, the aspects such as this external nutrition supply need to design for cell strain feature and metabolic characteristic, thus ensure higher culture density and taxol yield.The substratum of prior art is still not ideal enough in promotion yew cell secretion taxol.
Summary of the invention
The present invention is intended to the technological deficiency for prior art, there is provided a kind of Taxus chinensis cell cultures base promoting taxol to secrete, to solve in prior art the lower technical problem of the Taxus chinensis cell cultures base that promotes taxol the to secrete promoter action to its secretion taxol.
For realizing above technical purpose, the present invention by the following technical solutions:
A kind of Taxus chinensis cell cultures base promoting taxol to secrete, comprise the composition of following weight part: SODIUMNITRATE 8 ~ 10 parts, dipotassium hydrogen phosphate 4 ~ 6 parts, 2 ~ 3 parts, magnesium sulfate, 1 ~ 2 part, calcium chloride, peptone 1 ~ 2 part, Rsdix Wisterinae Venustae 1 ~ 2 part, Semen sesami Niger 1 ~ 2 part, Herba Lycopodii 1 ~ 2 part, Herba Caryopteridis Incanae 1 ~ 2 part, Gynura japonica 1 ~ 2 part, 50 ~ 60 parts, water.
As preferably, also comprise the invaluable of 1 ~ 2 weight part.
As preferably, also comprise the Rough Melic of 1 ~ 2 weight part.
As preferably, also comprise the Stigma Croci of 1 ~ 2 weight part.
As preferably, also comprise the Flower of Lalang Grass of 1 ~ 2 weight part.
As preferably, described substratum is grouped into by the one-tenth of following weight part: SODIUMNITRATE 9 parts, dipotassium hydrogen phosphate 5 parts, 2.5 parts, magnesium sulfate, 1.5 parts, calcium chloride, peptone 1.5 parts, Rsdix Wisterinae Venustae 1.5 parts, Semen sesami Niger 1.5 parts, Herba Lycopodii 1.5 parts, Herba Caryopteridis Incanae 1.5 parts, Gynura japonica 1.5 parts, 55 parts, water.
As preferably, described substratum is grouped into by the one-tenth of following weight part: SODIUMNITRATE 8.5 parts, dipotassium hydrogen phosphate 5.5 parts, 2.1 parts, magnesium sulfate, 1.3 parts, calcium chloride, peptone 1.7 parts, Rsdix Wisterinae Venustae 1.6 parts, Semen sesami Niger 1.2 parts, Herba Lycopodii 1.1 parts, Herba Caryopteridis Incanae 1.4 parts, Gynura japonica 1.5 parts, invaluable 1.4 parts, Rough Melic 1.1 parts, Stigma Croci 1.5 parts, Flower of Lalang Grass 1.8 parts, 57 parts, water.
In above technical scheme, described substratum is pulverizing after each composition mixing by formula ratio, is mixed to get with water, and described pulverizing can utilize all kinds of crushing material means well-known in the art to realize.Substratum of the present invention is particularly useful for the fluid suspension culture of yew cell, certainly also can be prepared as solid medium for seed culture or Secondary Culture.
The present invention by experiment means has extensively screened the nutritive substance as yew cell growth, determines formula of the present invention according to yew cell own metabolism characteristic and practical effect.This formula provides nitrogenous source and phosphorus source with traditional autotrophy substratum, and with the addition of certain calcium ion, and unexpected discovery helps lend some impetus to the productive rate of taxol by the Chinese medicinal materials adding particular types and composition on this basis.Based on discovery useful above, the present invention means are preferred by experiment Rsdix Wisterinae Venustae, Semen sesami Niger, Herba Lycopodii, Herba Caryopteridis Incanaes etc. are used for the Chinese medicinal materials composition of human disease treatment originally, gained substratum surprisingly obtains outstanding culture effect, the taxol yield of yew cell is significantly improved, and its production cycle is shorter simultaneously.The present invention obtains superior technique effect with the technique means of novelty, simultaneously cost lower, be easy to realize, therefore have a good application prospect.
Embodiment
Below will be described in detail the specific embodiment of the present invention.In order to avoid too much unnecessary details, in the examples below to belonging to known structure or function will not be described in detail.Apart from outside definition, technology used in following examples and scientific terminology have the identical meanings generally understood with those skilled in the art of the invention.
Embodiment 1
The Taxus chinensis cell cultures base promoting taxol to secrete, is grouped into by the one-tenth of following weight part: SODIUMNITRATE 8 parts, dipotassium hydrogen phosphate 4 parts, 2 parts, magnesium sulfate, 1 part, calcium chloride, peptone 1 part, Rsdix Wisterinae Venustae 1 part, Semen sesami Niger 1 part, Herba Lycopodii 1 part, Herba Caryopteridis Incanae 1 part, Gynura japonica 1 part, 50 parts, water.
Embodiment 2
The Taxus chinensis cell cultures base promoting taxol to secrete, is grouped into by the one-tenth of following weight part: SODIUMNITRATE 10 parts, dipotassium hydrogen phosphate 6 parts, 3 parts, magnesium sulfate, 2 parts, calcium chloride, peptone 2 parts, Rsdix Wisterinae Venustae 2 parts, Semen sesami Niger 2 parts, Herba Lycopodii 2 parts, Herba Caryopteridis Incanae 2 parts, Gynura japonica 2 parts, 60 parts, water.
Embodiment 3
A kind of Taxus chinensis cell cultures base promoting taxol to secrete, be grouped into by the one-tenth of following weight part: SODIUMNITRATE 9 parts, dipotassium hydrogen phosphate 5 parts, 2.3 parts, magnesium sulfate, 1.8 parts, calcium chloride, peptone 1.5 parts, Rsdix Wisterinae Venustae 1.6 parts, Semen sesami Niger 1.7 parts, Herba Lycopodii 1.8 parts, Herba Caryopteridis Incanae 1.2 parts, Gynura japonica 1.8 parts, 55 parts, water.
Embodiment 4
The Taxus chinensis cell cultures base promoting taxol to secrete, is grouped into by the one-tenth of following weight part: SODIUMNITRATE 8.8 parts, dipotassium hydrogen phosphate 5.4 parts, 2.7 parts, magnesium sulfate, 1.2 parts, calcium chloride, peptone 1.2 parts, Rsdix Wisterinae Venustae 1.2 parts, Semen sesami Niger 1.5 parts, Herba Lycopodii 1.6 parts, Herba Caryopteridis Incanae 1.4 parts, Gynura japonica 1.7 parts, invaluable 1 part, Flower of Lalang Grass 1.1 parts, 52 parts, water.
Embodiment 5
The Taxus chinensis cell cultures base promoting taxol to secrete, is grouped into by the one-tenth of following weight part: SODIUMNITRATE 9.4 parts, dipotassium hydrogen phosphate 4.2 parts, 2.1 parts, magnesium sulfate, 1.3 parts, calcium chloride, peptone 1.1 parts, Rsdix Wisterinae Venustae 1 part, Semen sesami Niger 2 parts, Herba Lycopodii 1.9 parts, Herba Caryopteridis Incanae 1.2 parts, Gynura japonica 1.9 parts, Rough Melic 1.3 parts, Stigma Croci 1.2 parts, Flower of Lalang Grass 1.6 parts, 58 parts, water.
Embodiment 6
The Taxus chinensis cell cultures base promoting taxol to secrete, is grouped into by the one-tenth of following weight part: SODIUMNITRATE 8.8 parts, dipotassium hydrogen phosphate 5.4 parts, 2.7 parts, magnesium sulfate, 1.2 parts, calcium chloride, peptone 1.2 parts, Rsdix Wisterinae Venustae 1.2 parts, Semen sesami Niger 1.5 parts, Herba Lycopodii 1.6 parts, Herba Caryopteridis Incanae 1.4 parts, Gynura japonica 1.7 parts, invaluable 1.5 parts, 55 parts, water.
Embodiment 7
The Taxus chinensis cell cultures base promoting taxol to secrete, is grouped into by the one-tenth of following weight part: SODIUMNITRATE 9.2 parts, dipotassium hydrogen phosphate 5.7 parts, 2.4 parts, magnesium sulfate, 1.5 parts, calcium chloride, peptone 1.5 parts, Rsdix Wisterinae Venustae 1.7 parts, Semen sesami Niger 1.7 parts, Herba Lycopodii 1.2 parts, Herba Caryopteridis Incanae 1.9 parts, Gynura japonica 2 parts, Stigma Croci 1.8 parts, 59 parts, water.
Embodiment 8
The Taxus chinensis cell cultures base promoting taxol to secrete, is grouped into by the one-tenth of following weight part: SODIUMNITRATE 8.5 parts, dipotassium hydrogen phosphate 5.5 parts, 2.1 parts, magnesium sulfate, 1.3 parts, calcium chloride, peptone 1.7 parts, Rsdix Wisterinae Venustae 1.6 parts, Semen sesami Niger 1.2 parts, Herba Lycopodii 1.1 parts, Herba Caryopteridis Incanae 1.4 parts, Gynura japonica 1.5 parts, invaluable 1.4 parts, Rough Melic 1.1 parts, Stigma Croci 1.5 parts, Flower of Lalang Grass 1.8 parts, 57 parts, water.
Above embodiments of the invention have been described in detail, but described content is only preferred embodiment of the present invention, not in order to limit the present invention.All make in application range of the present invention any amendment, equivalent to replace and improvement etc., all should be included within protection scope of the present invention.
Claims (7)
1. the Taxus chinensis cell cultures base promoting taxol to secrete, it is characterized in that the composition comprising following weight part: SODIUMNITRATE 8 ~ 10 parts, dipotassium hydrogen phosphate 4 ~ 6 parts, 2 ~ 3 parts, magnesium sulfate, 1 ~ 2 part, calcium chloride, peptone 1 ~ 2 part, Rsdix Wisterinae Venustae 1 ~ 2 part, Semen sesami Niger 1 ~ 2 part, Herba Lycopodii 1 ~ 2 part, Herba Caryopteridis Incanae 1 ~ 2 part, Gynura japonica 1 ~ 2 part, 50 ~ 60 parts, water.
2. the Taxus chinensis cell cultures base of promotion taxol secretion according to claim 1, characterized by further comprising the invaluable of 1 ~ 2 weight part.
3. the Taxus chinensis cell cultures base of promotion taxol secretion according to claim 1, characterized by further comprising the Rough Melic of 1 ~ 2 weight part.
4. the Taxus chinensis cell cultures base of promotion taxol secretion according to claim 1, characterized by further comprising the Stigma Croci of 1 ~ 2 weight part.
5. the Taxus chinensis cell cultures base of promotion taxol secretion according to claim 1, characterized by further comprising the Flower of Lalang Grass of 1 ~ 2 weight part.
6. the Taxus chinensis cell cultures base of promotion taxol secretion according to claim 1, it is characterized in that described substratum is grouped into by the one-tenth of following weight part: SODIUMNITRATE 9 parts, dipotassium hydrogen phosphate 5 parts, 2.5 parts, magnesium sulfate, 1.5 parts, calcium chloride, peptone 1.5 parts, Rsdix Wisterinae Venustae 1.5 parts, Semen sesami Niger 1.5 parts, Herba Lycopodii 1.5 parts, Herba Caryopteridis Incanae 1.5 parts, Gynura japonica 1.5 parts, 55 parts, water.
7. the Taxus chinensis cell cultures base of promotion taxol secretion according to claim 1, is characterized in that described substratum is grouped into by the one-tenth of following weight part: SODIUMNITRATE 8.5 parts, dipotassium hydrogen phosphate 5.5 parts, 2.1 parts, magnesium sulfate, 1.3 parts, calcium chloride, peptone 1.7 parts, Rsdix Wisterinae Venustae 1.6 parts, Semen sesami Niger 1.2 parts, Herba Lycopodii 1.1 parts, Herba Caryopteridis Incanae 1.4 parts, Gynura japonica 1.5 parts, invaluable 1.4 parts, Rough Melic 1.1 parts, Stigma Croci 1.5 parts, Flower of Lalang Grass 1.8 parts, 57 parts, water.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201510964059.7A CN105400731A (en) | 2015-12-18 | 2015-12-18 | Taxus chinensis cell culture medium promoting paclitaxel excretion |
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| CN201510964059.7A CN105400731A (en) | 2015-12-18 | 2015-12-18 | Taxus chinensis cell culture medium promoting paclitaxel excretion |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101319199A (en) * | 2007-06-08 | 2008-12-10 | 中国科学院大连化学物理研究所 | Method for cell abduction generation of taxone with Chinese yew |
| CN102187782A (en) * | 2010-03-10 | 2011-09-21 | 德阳市食用菌专家大院 | Method for culturing lucid ganoderma by using culture medium containing non-medicinal parts of Chinese medicinal materials |
-
2015
- 2015-12-18 CN CN201510964059.7A patent/CN105400731A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101319199A (en) * | 2007-06-08 | 2008-12-10 | 中国科学院大连化学物理研究所 | Method for cell abduction generation of taxone with Chinese yew |
| CN102187782A (en) * | 2010-03-10 | 2011-09-21 | 德阳市食用菌专家大院 | Method for culturing lucid ganoderma by using culture medium containing non-medicinal parts of Chinese medicinal materials |
Non-Patent Citations (2)
| Title |
|---|
| 周忠强等: "红豆杉细胞培养生产紫杉醇的研究进展", 《中南民族大学学报》 * |
| 李永成: "东北红豆杉悬浮细胞与内生真菌在紫杉醇合成中相互关系的研究", 《中国博士学位论文全文数据库农业科技辑》 * |
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