CN107126553A - A kind of Endometrial stem cell preparation and its application - Google Patents
A kind of Endometrial stem cell preparation and its application Download PDFInfo
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- CN107126553A CN107126553A CN201710306817.5A CN201710306817A CN107126553A CN 107126553 A CN107126553 A CN 107126553A CN 201710306817 A CN201710306817 A CN 201710306817A CN 107126553 A CN107126553 A CN 107126553A
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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
- A61K38/385—Serum albumin
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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
- A61K35/48—Reproductive organs
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- A—HUMAN NECESSITIES
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- 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
- A61K38/18—Growth factors; Growth regulators
- A61K38/1858—Platelet-derived growth factor [PDGF]
- A61K38/1866—Vascular endothelial growth factor [VEGF]
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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
- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/20—Interleukins [IL]
- A61K38/2066—IL-10
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Abstract
The application belongs to regenerative medicine and biology techniques field, and in particular to a kind of Endometrial stem cell preparation and its application.Endometrial stem cell preparation provided by the present invention is included：Endometrial stem cell, human serum albumins, IL-10 and vascular endothelial growth factor, source of human stem cell, convenient material drawing, non-invasive, no ethics limitation are used as from Endometrial stem cell；Endometrial stem cell and cell factor synergy, have effectively facilitated the reparation of uterine cicatrix, and improve the conversion ratio of stem cell medicine.Present invention also offers application of the above-mentioned Endometrial stem cell preparation in uterine cicatrix medicine is prepared.
Description
Technical field
The invention belongs to regenerative medicine and biology techniques field, and in particular to a kind of Endometrial stem cell preparation and its
Using.
Background technology
In recent years, induced abortion, induced abortion patients number increase year by year in clinic, and multiple induced abortion is easy in a short time
Cause the damage of endometrium so that endometrial growth difference and Asherman's syndrom, add endometrium secondary inflammation
Risk so that the risk of secondary infertility significantly increases.At present, the stem cell medicine applied to uterine cicatrix reparation mainly has
Bone marrow stem cell preparation and autologous fat stem cell preparation, its source of human stem cell are required to adopt by wound progress sample
Collection, not only samples difficulty, and certain somatic damage is easily caused to patient.
The content of the invention
In view of this, it is sub applied to scar is prepared it is an object of the invention to provide a kind of Endometrial stem cell preparation
Palace medicine, source of human stem cell is convenient, non-invasive.
The concrete technical scheme of the present invention is as follows：
The invention provides a kind of Endometrial stem cell preparation, comprising：Endometrial stem cell, human serum albumins,
IL-10 and vascular endothelial growth factor.
Endometrial stem cell is a kind of adult stem cell, the potential with self-renewing, Multidirectional Differentiation and hyperproliferation,
Carcinogenic risk is low, is disputed on by ethics few.Therefore, the related researcher of this area thinks that Endometrial stem cell is likely to become
Endometrium is excessively thin and intimal lesion patient carries out inner membrance and repairs replacement therapy means.In view of Endometrial stem cell and intrauterine
The substantial connection of film growth, from Isolation and culture Endometrial stem cell, is then transplanted in endometrium and promotes uterus scar
Trace is repaired.
It is preferred that, the cell concentration of the Endometrial stem cell is 5.0 × 106~2.0 × 107Individual/mL.
It is preferred that, the concentration of volume percent of the human serum albumins is 1%~10%, preferably 5%.
The participation of a variety of growth factors is needed in uterine cicatrix repair process, but Endogenous Growth Factors are often difficult to
Needs are met, therefore, timely and appropriately introducing specific exogenous growth factor contributes to the reparation of uterus scar, by screening
Optimization, the exogenous growth factor is stem cell factor and stroma cell derivative factor.Wherein, IL-10 (IL-
10) it is a kind of many cells source, multi-functional cell factor, adjusts the growth and differentiation of cell, participates in inflammatory reaction and immune anti-
Should, it is generally acknowledged at present inflammation and immunosuppressive factor；Vascular endothelial growth factor (VEGF) is specific effect in interior
The glycosylation cell mitogen element of chrotoplast, permeability, induction of vascular in enhancing blood vessel occur and angiogenesis and endothelium
Played a role in terms of cell growth, promotion cell migration, suppression Apoptosis.
It is preferred that, the concentration of the IL-10 is 5~30ng/mL.
It is preferred that, the concentration of the vascular endothelial growth factor is 5~30ng/mL.
It is preferred that, the Endometrial stem cell preparation also includes：Solvent, the solvent is physiological saline.
Present invention also offers a kind of preparation method of above-mentioned Endometrial stem cell preparation, by Endometrial stem cell,
Human serum albumins, IL-10, vascular endothelial growth factor and physiological saline mixing, obtain the endometrium dry thin
Born of the same parents' preparation.
The endometrium obtained present invention also offers above-mentioned Endometrial stem cell preparation or above-mentioned preparation method is dry thin
Application of born of the same parents' preparation in uterine cicatrix medicine is prepared.
Compared with prior art, Endometrial stem cell preparation of the invention has advantages below：
1) present invention is used as source of human stem cell, convenient material drawing, non-invasive, no ethics limit from Endometrial stem cell
System；It is safe and reliable and the Endometrial stem cell is autologous Endometrial stem cell, can avoid immunological rejection；
2) Endometrial stem cell preparation of the present invention also includes IL-10 and vascular endothelial growth factor, and both assist
Same-action, improves stem cell transformation rate, promotes the reparation of uterine cicatrix；
3) invention formulation is from physiological saline as solvent, and its osmotic pressure is consistent with the osmotic pressure of human body, can avoid damaging
Hinder its bioactivity under uterine tissue, Cord blood stable；
4) preparation process of invention formulation is simple, takes short, achievable large-scale production.
Brief description of the drawings
In order to illustrate more clearly about the embodiment of the present invention or technical scheme of the prior art, below will be to embodiment or existing
There is the accompanying drawing used required in technology description to be briefly described, it should be apparent that, drawings in the following description are only this
The embodiment of invention, for those of ordinary skill in the art, on the premise of not paying creative work, can also basis
The accompanying drawing of offer obtains other accompanying drawings.
Fig. 1 is the microphoto of second generation Endometrial stem cell；Wherein, Fig. 1 a are that second generation Endometrial stem cell is put
Big 40 times of microphoto, Fig. 1 b are the microphoto that second generation Endometrial stem cell amplifies 100 times；
Fig. 2 is the qualification result of the surface antigen feature of Endometrial stem cell；
Fig. 3 is the result for the stem cell medicine Endometrial stem cell vitality test that embodiment 2 is obtained.
Embodiment
By being used as core cell from Endometrial stem cell replacement stem cell and fat cell, while optimizing system
Constituent in agent, the invention provides a kind of Endometrial stem cell preparation, applied to uterine cicatrix medicine is prepared, does thin
Born of the same parents' survival rate stabilization, convenient sources, non-invasive, and the preparation time of preparation is short, efficiency high.
The present invention is to Endometrial stem cell, human serum albumins, IL-10 and vascular endothelial growth factor
Source is not particularly limited, using Endometrial stem cell well-known to those skilled in the art, human serum albumins, interleukin
10 and vascular endothelial growth factor, or prepared using routine techniques well-known to those skilled in the art
Endometrial stem cell, human serum albumins, IL-10 and vascular endothelial growth factor.
Technical scheme is clearly and completely described below in conjunction with the specific embodiment of the invention, it is clear that
Described embodiment is a part of embodiment of the invention, rather than whole embodiments.Those skilled in the art should manage
Solution, modifies to the specific embodiment of the present invention or some technical characteristics is replaced on an equal basis, without departing from the present invention
The spirit of technical scheme, all should cover in the scope of protection of the invention.
Human serum albumins in following examples is purchased from go the same way bio-pharmaceuticals limited company, IL-10 and blood vessel
Endothelial growth factor is purchased from Shanghai Hua Ying biological medicines Science and Technology Ltd..
The preparation of the Endometrial stem cell of embodiment 1
After agreeing to through consulting, received according to menses collection suit (Hangzhou Yi Wensai, S-Evans biosciences) specification
Collect healthy women through blood sample；Then these samples are transferred to the phosphoric acid containing dual anti-(penicillin/streptomycin) and heparin
In salt buffer, preserved under the conditions of 4 DEG C, sample is sent to laboratory centrifugal within 24h~48h；Then supernatant is taken and in nothing
Quick detection whether there is germ contamination under the conditions of bacterium, if feminine gender then continues to be separately cultured and identify, Endometrial stem cell
Be separately cultured and qualification process refer to following documents：
Zhou Hua, Song Lina, quick people's Endometrial stem cell in-vitro separation, culture and the Chinese traditional Chinese medicine academic periodicals of identification
[J].2014.32(8):1907-1908。
The preparation and its evaluation of the Endometrial stem cell preparation of embodiment 2
1st, a kind of preparation of Endometrial stem cell preparation：
1) human serum albumin solution is obtained using normal saline dilution human serum albumins, wherein human serum albumins
Concentration of volume percent is 5%；
2) toward step 1) in add IL-10 and VEGF respectively in obtained human serum albumin solution so that IL-10 end
Concentration is 10ng/mL, VEGF final concentration of 30ng/mL；
3) using step 2) the obtained human serum albumin solution containing IL-10, VEGF is resuspended what is obtained in embodiment 1
Endometrial stem cell, obtains the Endometrial stem cell preparation, the wherein concentration of Endometrial stem cell is 5.0 × 106
Individual/mL；
4) step 3) obtained Endometrial stem cell dispenses into syringe, is then put into sterile sylphon,
The sylphon for turning to have preparation is transferred in the storage box for being placed with ice bag, is transported at 0~4 DEG C.
2nd, effect of Endometrial stem cell preparation and quality evaluation
1) method for building up of animal model refers to documents below：
Xu builds ripple intraperitoneal injections and sets up C57BL/6 endometrium of mouse Endometriosis models, University Of Suzhou's master's degree opinion
Text.
2) evaluation criterion
A. imaging observation (x-ray scoring)；
B. histology (different time points new bone area percent).
3) cell and quality of the pharmaceutical preparations evaluation
A. cell quality evaluation (form, surface antigen, vigor)
Fig. 1 is the microphoto of second generation Endometrial stem cell, as illustrated, the state of Endometrial stem cell is good
Good, multiplication capacity is strong, and substantially, bag slurry is full for cell refractive power, and cell grows in fusiformis, meets the form of mescenchymal stem cell；Fig. 2
For using qualification result of the flow cytometer to Endometrial stem cell surface antigen feature, as shown in the results, the high table of the cell
Up to CD59 and CD90, expression rate is respectively 99.4%, 99.5%；Low expression HLA-DR and CD45, expression rate are respectively 0.5% He
0.4%, meet the surface antigen feature of mescenchymal stem cell, illustrate that Endometrial stem cell does not occur the phenomenon of differentiation,
The characteristics of still maintain stem cell；Fig. 3 is that the Endometrial stem cell vigor in stem cell medicine is detected using Trypan Blue
Result, as illustrated, cellular morphology is small and homogeneous, refractivity is strong, and most cells are living cells, and a small amount of cell is dead thin
Born of the same parents, carry out three survival rate tests, its Cell viability is respectively 94.67%, 95.16%, 94.13%.
B. quality of the pharmaceutical preparations evaluation (bacterium, fungi, endotoxin, five viroids)
By detection, Endometrial stem cell preparation is in bacterium, fungi, endotoxin and five big viral (hepatitis A, hepatitis B, third
Liver, syphilis and HIV) detection in, be as a result negative.The up-to-standard of Endometrial stem cell preparation is illustrated, be can be used for
Re-injection ejection preparation in body, promotes the reparation of uterine cicatrix.
Embodiment 3 investigates Healing of the different dry cell content to uterine cicatrix
Stem cell medicine is prepared respectively by formula table 1 Suo Shi, then carries out x-ray scoring, its result is as shown in table 2.Lane-
The scoring display of Sandhu x-rays：Postoperative 2nd, 4,8 weeks, A groups have statistical significance (P compared with control group 1, not>0.05)；B groups,
The uterus scar repairing effect of C in combination D groups is substantially better than control group 1 and A groups, and difference has statistical significance (P<0.05), and
And D composition bones are better than B groups and C groups, the statistically significant (P of difference<0.05).As a result show that the cell worked as in stem cell medicine is dense
Spend for 5.0 × 106~2.0 × 107During individual/mL, the effect that it repairs uterus scar is more obvious.
Table 1
| Group | Concrete composition |
| Control group 1 | 5% human serum albumins |
| A groups | 2.5×106Individual/human serum albumins of mL Endometrial stem cells+5% |
| B groups | 5.0×106Individual/human serum albumins of mL Endometrial stem cells+5% |
| C groups | 1.0×107Individual/human serum albumins of mL Endometrial stem cells+5% |
| D groups | 2.0×107Individual/human serum albumins of mL Endometrial stem cells+5% |
The scoring of the postoperative different time points x-ray of table 2 compare (n=3,)
* represent compared with control group, P<0.05, with statistical significance.
Embodiment 4
Stem cell medicine is prepared respectively by formula table 3 Suo Shi, is then carried out x-ray scoring and is calculated the repair surface of uterine cicatrix
Product percentage, its result is as shown in 3~table of table 5.
Lane-Sandhu x-rays are scored and uterine cicatrix repaired area percentage comparing result is shown：Postoperative 2nd, 4,8
In week, E groups~G groups are compared with control group 1, control group 2, and difference has statistical significance (P<0.05)；H groups and control group 1, compare
Group 2 is compared, and difference has extremely significant statistical significance (P<0.01).
Table 3
The scoring of the postoperative different time points x-ray of table 4 compare (n=8,)
* represent compared with control group, P<0.05, with statistical significance；
# represented compared with control group, P<0.01, with extremely significant statistical significance.
The postoperative different time points uterus scar repairing area percentage of table 5
| Group | Number of elements | 2 weeks | 4 weeks | 8 weeks |
| Control group 1 | 8 | 6.9±1.5 | 14.8±2.9 | 23.6±4.7 |
| Control group 2 | 8 | 8.4±2.7 | 20.4±3.1 | 32.1±2.9 |
| E groups | 8 | 11.4±4.8* | 23.7±3.6* | 40.8±4.2* |
| F groups | 8 | 20.6±2.8* | 35.7±3.9* | 48.3±6.5* |
| G groups | 8 | 21.2±3.4* | 36.6±4.1* | 47.4±2.8* |
| H groups | 8 | 28.5±4.7# | 46.8±6.5# | 60.2±5.4# |
* represent compared with control group, P<0.05, with statistical significance；
# represented compared with control group, P<0.01, with extremely significant statistical significance.
Claims (8)
1. a kind of Endometrial stem cell preparation, comprising：Endometrial stem cell, human serum albumins, IL-10 and blood vessel
Endothelial growth factor.
2. Endometrial stem cell preparation according to claim 1, it is characterised in that the Endometrial stem cell it is thin
Born of the same parents' concentration is 5.0 × 106~2.0 × 107Individual/mL.
3. Endometrial stem cell preparation according to claim 1, it is characterised in that the volume of the human serum albumins
Percent concentration is 1%~10%.
4. Endometrial stem cell preparation according to claim 1, it is characterised in that the concentration of the IL-10 is 5
~30ng/mL.
5. Endometrial stem cell preparation according to claim 1, it is characterised in that the vascular endothelial cell growth because
The concentration of son is 5~30ng/mL.
6. Endometrial stem cell preparation according to claim 1, it is characterised in that also include：Solvent, the solvent is
Physiological saline.
7. the preparation method of the Endometrial stem cell preparation described in a kind of claim 6, it is characterised in that do endometrium
Cell, human serum albumins, IL-10, vascular endothelial growth factor and physiological saline mixing, obtain the intrauterine
Film stem cell medicine.
8. preparation method as described in the Endometrial stem cell preparation or claim 7 as described in claim 1 to 6 any one is obtained
To Endometrial stem cell preparation prepare uterine cicatrix medicine in application.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113181218A (en) * | 2021-03-22 | 2021-07-30 | 中国福利会国际和平妇幼保健院 | Application of human amniotic epithelial cells in preparation of preparation for repairing uterine scar cells |
| CN115300612A (en) * | 2022-07-29 | 2022-11-08 | 海南博鳌未来医院有限公司 | Stem cell preparation for repairing endometrium and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105687240A (en) * | 2016-03-14 | 2016-06-22 | 广州赛莱拉干细胞科技股份有限公司 | Menstrual blood source stem cell preparation and preparation method and application thereof |
| CN106267161A (en) * | 2016-09-30 | 2017-01-04 | 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 |
-
2017
- 2017-04-27 CN CN201710306817.5A patent/CN107126553A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105687240A (en) * | 2016-03-14 | 2016-06-22 | 广州赛莱拉干细胞科技股份有限公司 | Menstrual blood source stem cell preparation and preparation method and application thereof |
| CN106267161A (en) * | 2016-09-30 | 2017-01-04 | 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 |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113181218A (en) * | 2021-03-22 | 2021-07-30 | 中国福利会国际和平妇幼保健院 | Application of human amniotic epithelial cells in preparation of preparation for repairing uterine scar cells |
| CN115300612A (en) * | 2022-07-29 | 2022-11-08 | 海南博鳌未来医院有限公司 | Stem cell preparation for repairing endometrium and application thereof |
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