CN106492197A - A kind of stem cell medicine and its preparation method and application - Google Patents
A kind of stem cell medicine and its preparation method and application Download PDFInfo
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- CN106492197A CN106492197A CN201611086967.1A CN201611086967A CN106492197A CN 106492197 A CN106492197 A CN 106492197A CN 201611086967 A CN201611086967 A CN 201611086967A CN 106492197 A CN106492197 A CN 106492197A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/25—Araliaceae (Ginseng family), e.g. ivy, aralia, schefflera or tetrapanax
- A61K36/258—Panax (ginseng)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/38—Albumins
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Abstract
The present invention relates to biological therapy field, discloses a kind of stem cell medicine and its preparation method and application, the stem cell medicine has good curative effect to hepatic injury.Stem cell medicine disclosed by the invention, including：Mescenchymal stem cell, albumin, ginsenoside and solvent.The invention discloses the preparation method of the stem cell medicine, comprises the following steps：A), the mixing of mescenchymal stem cell, albumin, ginsenoside and solvent, obtains stem cell medicine.The invention also discloses the application of the stem cell medicine or stem cell medicine obtained in the preparation method in treatment liver injury medicament is prepared.
Description
Technical field
The present invention relates to biological therapy field, more particularly to a kind of stem cell medicine and its preparation method and application.
Background technology
Mescenchymal stem cell is one kind of multipotential stem cell, and the potential with Multidirectional Differentiation, under conditions of inducing in vitro
Various Tissues and the cells such as muscle, nerve, bone, cartilage, fat, cardiac muscle and class hepatic tissue can be divided into.After Long Term Passages
Still there is propagation replication capacity, is a kind of seed cell of preferable histoorgan engineering.Clinically also have at present more next
More people are applied to reparation and the immune system of histoorgan using mescenchymal stem cell.
Prior art using intravenous injection culture mescenchymal stem cell treating acute liver damage, the shortcoming of prior art
That to depend merely on mescenchymal stem cell treatment hepatic injury effect poor because different patient the reaction of mescenchymal stem cell is had aobvious
Difference is write, it is the cell through cultivating to add mescenchymal stem cell, it is easy to disposed by body.Therefore, study a kind of to liver
It is that those skilled in the art need the technology for solving to ask to damage the mescenchymal stem cell treatment method with good efficacy or preparation
Topic.
Content of the invention
In view of this, the invention discloses a kind of stem cell medicine and its preparation method and application, the stem cell medicine
There is good curative effect to hepatic injury.
The invention discloses a kind of stem cell medicine, including：Mescenchymal stem cell, albumin, ginsenoside and solvent.
In some specific embodiments of the present invention, the content of the mescenchymal stem cell is 3 × 105～5 × 106/
ml.Preferably, the content of the mescenchymal stem cell is 1 × 106Individual/ml.
In some specific embodiments of the present invention, the albuminous content is 1%～10% (w/v).Preferably,
The albuminous content is 3%.
In some specific embodiments of the present invention, the content of the ginsenoside is 20ng/ml～120ng/ml.Excellent
Selection of land, the content of the ginsenoside is 60ng/ml.
Preferably, the solvent is physiological saline.
The invention also discloses the preparation method of the stem cell medicine, comprises the following steps：
A), the mixing of mescenchymal stem cell, albumin, ginsenoside and solvent, obtains stem cell medicine.
Preferably, the mescenchymal stem cell is obtained according to following steps：
B1), marrow is centrifuged, primary mescenchymal stem cell is obtained；
B2), the resuspended primary mescenchymal stem cell, is cultivated；
B3), change liquid and remove non-attached cell, add culture medium to continue to cultivate the primary mescenchymal stem cell,
The culture medium includes ganoderma lucidum triterpene compounds；
B4), the cell to cultivating digests, and obtains mescenchymal stem cell.
The invention also discloses the stem cell medicine or stem cell medicine obtained in the preparation method are preparing treatment
Application in liver injury medicament.
The present invention passes through to add ganoderma lucidum triterpene compounds in mescenchymal stem cell culture, and adds in stem cell medicine
Enter ginsenoside, can significantly inhibit the immune response in patient's body, and strengthen the repair ability of mescenchymal stem cell, improve
Therapeutic effect.
In sum, the invention discloses a kind of stem cell medicine and its preparation method and application, the stem cell medicine can
To suppress the immune response in patient's body, mescenchymal stem cell is made from the attack of patient's vivo immuning system, increase cell and exist
Internal life span, and there is good curative effect to hepatic injury.
Description of the drawings
Fig. 1 shows the results of serological detection of experimental group, control group and comparative example group after 1 week；
Fig. 2 shows the results of serological detection of experimental group, control group and comparative example group after 4 weeks；
Fig. 3 shows the results of serological detection of experimental group, control group and comparative example group after 8 weeks.
Specific embodiment
The invention discloses a kind of stem cell medicine and its preparation method and application, the stem cell medicine can suppress patient's body
Interior immune response, has good curative effect to hepatic injury.Those skilled in the art can use for reference present disclosure, be suitably modified work
Skill parameter is realized.Specifically, all similar replacements and change are aobvious and easy for a person skilled in the art
See, they are considered as being included in the present invention.The method of the present invention and application are described by preferred embodiment,
Related personnel substantially method described herein and application can be modified in without departing from present invention, spirit and scope or
Suitably change and combine, realize and apply the technology of the present invention.
In the present invention, involved raw material can be buied by market.Wherein, from Zhengzhou Lay scholar, ginsenoside is purchased for albumin purchase
Buy from Shanghai Aladdin biochemical technology limited company, ganoderma lucidum triterpene compounds purchase is limited from Dalian U.S. logical sequence biotechnology
Company.
With reference to embodiment, the present invention is expanded on further.
The preparation being separately cultured with stem cell medicine of 1 mescenchymal stem cell of embodiment
1st, mouse bone marrow cells 10ml that aseptic condition under extract are taken, after adding isopyknic DMEM-LG culture mediums dilution to mix,
1000r/min is centrifuged 5min, removes the fat and supernatant on upper strata；
2nd, with the DMEM-F12 culture medium re-suspended cells containing 10%FBS and 1 × dual anti-, and adjust cell density for 5 ×
105/ ml is inoculated in 15cm culture dishes, is transferred to 5%CO2, 37 DEG C, the CO that saturated humidity is 95%2Cultivate in incubator；
3rd, liquid is changed after culture 48h and remove not adherent heteroproteose cell, while adding the ganoderma lucidum triterpene of 50ng/ml in culture medium
Compound, hereafter changes liquid 1 time, the same ganoderma lucidum triterpene compounds for adding 50ng/ml every 2～3d；
4th, treat that cell number reaches 1 × 108Culture medium is discarded after individual, and physiological saline is cleaned 3 times, adds 8ml0.25% (quality
Volume ratio) pancreatin (purchase from gibco companies) vitellophag；
5th, after cell rounding loosening, the DMEM-LG culture mediums containing 10%FBS of twice pancreatin volume are added to terminate disappearing
Change；
6th, pour out cell suspension, be centrifuged supernatant discarded, physiological saline is cleaned 2 times, obtain for preparing stem cell medicine
Mescenchymal stem cell (CD90>95%, CD105>95%, CD74>95%, CD34<2%, CD45<2%, HLADR<2%).
7th, with the resuspended obtained mescenchymal stem cell of the physiological saline containing 3% albumin and 60ng/ml ginsenosides, make
Cell density is 1 × 106Individual/ml, makes stem cell medicine parenteral solution.
The preparation being separately cultured with stem cell medicine of comparative example mescenchymal stem cell
1st, mouse bone marrow cells 10ml that aseptic condition under extract are taken, after adding the dilution of DMEM-LG culture mediums equimultiple to mix,
1000r/min is centrifuged 5min, removes the fat and supernatant on upper strata；
2nd, with the DMEM-F12 culture medium re-suspended cells containing 10%FBS and 1 × dual anti-, and adjust cell density for 5 ×
105/ ml is inoculated into 15cm culture dishes, is transferred to 5%CO2, 37 DEG C, the CO that saturated humidity is 95%2Cultivate in incubator.
3rd, liquid is changed after culture 48h and remove not adherent heteroproteose cell, hereafter change liquid 1 time every 2～3d；
4th, treat that cell number reaches 1 × 108Culture medium is discarded after cell, physiological saline is cleaned 3 times, add the digestion of 8ml pancreatin
Cell；
5th, after cell rounding loosening, the DMEM-LG culture mediums containing 10%FBS of twice pancreatin volume are added to terminate disappearing
Change.
6th, pour out cell suspension, be centrifuged supernatant discarded, physiological saline is cleaned 2 times, obtain for preparing stem cell medicine
Mescenchymal stem cell；
7th, with the resuspended mescenchymal stem cell of the physiological saline containing 3% human serum albumin, it is 1 × 10 to make cell density6Individual/
Ml, makes parenteral solution.
3 animal experiment of embodiment
Purchase is divided into three groups from 12 kunming mices of the acute hepatic injury model at Guangdong Province medical animal experiment center,
Group 1 is test group, injects stem cell medicine prepared by embodiment 1；Group 2 is the physiology salt of control group, injection and test group equivalent
Water；Group 3 is comparative example group, injects stem cell medicine prepared by comparative example.Injection system is tail vein injection, once a day,
Volume injected is 3ml, continuous injection 3 days.
Corresponding data is detected behind 1 week, 4 weeks, 8 weeks.
1 week after the treatment, 4 weeks, 8 weeks per group randomly selecting 2 mouse, blood drawing by Elisa kits detect AST,
ALT indexs (kit is bought from Nanjing Jin Yibai bio tech ltd).
After 1 week, results of serological detection is shown in Table 1, Fig. 1；After 4 weeks, results of serological detection is shown in Table 2, Fig. 2；Serology after 8 weeks
Testing result is shown in Table 3, Fig. 3.
Table 1
ALT and AST be detect liver function two indexs, the normal range (NR) of numerical value between 0 and 40, in liver function damage
In the case of numerical value can raise, and numerical value is higher, shows that liver function damage is more serious.Experimental group can be drawn from data
Liver function level is better than control combination comparative example group, and comparative example group is better than control group again, with significant difference (P ＜
0.05).
Table 2
Can show that from data the liver function level of experimental group is better than and compare combination comparative example group, and comparative example group
Control group is better than, with significant difference (P ＜ 0.05).
Table 3
Can show that from data the liver function level of experimental group is better than and compare combination comparative example group, and comparative example group
Control group is better than, with significant difference (P ＜ 0.05).
The above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (8)
1. a kind of stem cell medicine, it is characterised in that include：Mescenchymal stem cell, albumin, ginsenoside and solvent.
2. stem cell medicine according to claim 1, it is characterised in that the content of the mescenchymal stem cell is 3 × 105
～5 × 106/ml.
3. stem cell medicine according to claim 1, it is characterised in that the albuminous content is 1%～10% (w/
v).
4. stem cell medicine according to claim 1, it is characterised in that the content of the ginsenoside be 20ng/ml～
120ng/ml.
5. stem cell medicine according to claim 1, it is characterised in that the solvent is physiological saline.
6. the preparation method of the stem cell medicine described in claim 1 to 5 any one, it is characterised in that take mesenchyma dry thin
The mixing of born of the same parents, albumin, ginsenoside and solvent, obtains stem cell medicine.
7. preparation method according to claim 6, the mescenchymal stem cell are obtained according to following steps：
B1), marrow is centrifuged, primary mescenchymal stem cell is obtained；
B2), the resuspended primary mescenchymal stem cell, is cultivated；
B3), change liquid and remove non-attached cell, add culture medium to continue to cultivate the primary mescenchymal stem cell, described
Culture medium includes ganoderma lucidum triterpene compounds；
B4), the cell to cultivating digests, and obtains mescenchymal stem cell.
8. the stem cell medicine described in Claims 1 to 5 or stem cell system obtained in the preparation method described in claim 6 or 7
Application of the agent in treatment liver injury medicament is prepared.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106729642A (en) * | 2017-03-17 | 2017-05-31 | 广州赛莱拉干细胞科技股份有限公司 | A kind of stem cell medicine and its preparation method and application |
| CN114515296A (en) * | 2022-02-25 | 2022-05-20 | 和携科技有限公司 | Preparation method of adipose-derived mesenchymal stem cell secretory factor |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102028970A (en) * | 2010-12-27 | 2011-04-27 | 协和干细胞基因工程有限公司 | Stem cell preparation for treating cirrhosis |
| CN102949418A (en) * | 2012-06-02 | 2013-03-06 | 郑家德 | Composition of human stem cells and ganoderma spore biological activity substances and preparation method thereof |
| CN103045531A (en) * | 2011-10-14 | 2013-04-17 | 北京清美联创干细胞科技有限公司 | Application of ginsenoside Rd for inducing mesenchymal stem cell to be directionally differentiated in to hepatic cell in vitro |
| CN106215171A (en) * | 2016-09-30 | 2016-12-14 | 广州赛莱拉干细胞科技股份有限公司 | A kind of mesenchymal stem cell injection and its preparation method and application |
-
2016
- 2016-11-30 CN CN201611086967.1A patent/CN106492197A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102028970A (en) * | 2010-12-27 | 2011-04-27 | 协和干细胞基因工程有限公司 | Stem cell preparation for treating cirrhosis |
| CN103045531A (en) * | 2011-10-14 | 2013-04-17 | 北京清美联创干细胞科技有限公司 | Application of ginsenoside Rd for inducing mesenchymal stem cell to be directionally differentiated in to hepatic cell in vitro |
| CN102949418A (en) * | 2012-06-02 | 2013-03-06 | 郑家德 | Composition of human stem cells and ganoderma spore biological activity substances and preparation method thereof |
| CN106215171A (en) * | 2016-09-30 | 2016-12-14 | 广州赛莱拉干细胞科技股份有限公司 | A kind of mesenchymal stem cell injection and its preparation method and application |
Non-Patent Citations (2)
| Title |
|---|
| OWEN ANDREW等: "mesenchymal stromal cell therapy in liver disease：opportunities and lessons to be learnt", 《AMERICAN JOURNAL OF PHYSIOLOGY-GASTROINTESTINAL AND LIVER PHYSIOLOGY》 * |
| 程建文等: "芒果苷对缺氧损伤骨髓间充质干细胞的保护作用", 《中国组织工程》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106729642A (en) * | 2017-03-17 | 2017-05-31 | 广州赛莱拉干细胞科技股份有限公司 | A kind of stem cell medicine and its preparation method and application |
| CN114515296A (en) * | 2022-02-25 | 2022-05-20 | 和携科技有限公司 | Preparation method of adipose-derived mesenchymal stem cell secretory factor |
| CN114515296B (en) * | 2022-02-25 | 2024-01-12 | 和携科技有限公司 | Preparation method of adipose-derived mesenchymal stem cell secretion factor |
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Application publication date: 20170315 |