CN107384858A - A kind of preparation method and applications of hypoxic tolerance type mescenchymal stem cell - Google Patents
A kind of preparation method and applications of hypoxic tolerance type mescenchymal stem cell Download PDFInfo
- Publication number
- CN107384858A CN107384858A CN201710704337.4A CN201710704337A CN107384858A CN 107384858 A CN107384858 A CN 107384858A CN 201710704337 A CN201710704337 A CN 201710704337A CN 107384858 A CN107384858 A CN 107384858A
- Authority
- CN
- China
- Prior art keywords
- stem cell
- mescenchymal stem
- tolerance type
- cell
- hypoxic tolerance
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0662—Stem cells
- C12N5/0668—Mesenchymal stem cells from other natural sources
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2500/00—Specific components of cell culture medium
- C12N2500/02—Atmosphere, e.g. low oxygen conditions
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Developmental Biology & Embryology (AREA)
- Biomedical Technology (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Hematology (AREA)
- Rheumatology (AREA)
- Virology (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention discloses a kind of preparation method and applications of hypoxic tolerance type mescenchymal stem cell, the hypoxic tolerance type source for mesenchymal stem cells is in people's umbilical cord China Tong Shi glue, condition of culture is 1 6% oxygen concentration, compared with the mescenchymal stem cell cultivated under excess oxygen, hypoxic tolerance type mescenchymal stem cell has stronger paracrine function and survival ability；In animal model of acute myocardial infarction, hypoxic tolerance type mescenchymal stem cell can effectively facilitate angiogenesis, and can strengthen the power of regeneration of cardiac muscle cell；The hypoxic tolerance type mescenchymal stem cell of the present invention prepares simple, stability height, good effect, can be as the preferred scheme of clinical treatment acute myocardial infarction AMI.
Description
Technical field
The present invention relates to stem cells technology field, and in particular to a kind of preparation method of hypoxic tolerance type mescenchymal stem cell
And its application in acute myocardial infarction AMI is treated.
Background technology
Acute myocardial infarction AMI (acute myocardial infarction, AMI) is that one kind is closed because coronary artery is acute
Plug, causes blood supply drastically to reduce or interrupt and cause the disease of cardiac muscle cell's ischemic necrosis, belongs in angiocardiopathy
Common severe crisis, it is dead caused by acute myocardial infarction AMI to account for more than the 50% of angiocardiopathy death toll.Given birth to
The influence of the factors such as living mode, aging population, China's myocardial infarction incidence of disease is in obvious ascendant trend, estimate to the year two thousand thirty I
State's myocardial infarction patient quantity is up to 23,000,000.At present, the mode of clinical treatment acute myocardial infarction is mainly performed the operation (percutaneous to be preced with
Shape aortic procedures) and medicine (antithrombotic), both modes can prevent the expansion of myocardial infarct size, to a certain degree
The upper alleviation state of an illness, very weak yet with Human adult cardiomyocytes power of regeneration, having occurred and that the cardiac muscle of infarct can not effectively be repaiied
It is multiple.It by way of being transfused exogenous stem cells, can directly or indirectly promote cardiac muscle mitochondria, effectively repair what is be damaged
Cardiac muscle.Stem-cell therapy acute myocardial infarction AMI is of increased attention.
Mescenchymal stem cell (Mesenchymal Stem Cells, MSCs) derives from the mesoderm of mesoderm growing early stage, is one
Class has the initial cell of self-replacation and multi-lineage potential, is widely present in each tissue of body and organ, such as newborn
Youngster's umbilical cord, marrow, fat etc.；It can be divided into Various Tissues specific cell under given conditions, such as Gegenbaur's cell, cartilage
Cell, adipocyte, nerve cell, cardiac muscle cell etc..In terms of acute myocardial infarction AMI is treated, MSCs can move to pars affecta
Position is simultaneously activated, and so as to paracrine cytokine profiles, on the one hand can activate Cardiac Stem Cells, promotes it to breed and divide
Change；On the other hand angiogenesis can also be promoted, recovers local blood supply.Although MSCs acute myocardial infarction treatment in security and
Validity is confirmed, but stem-cell therapy is also faced with many difficulties and challenge.
The factors such as MSCs source, dosage and infusion have a significant impact for therapeutic effect, MSCs returning in vivo
Nest effect and time-to-live are also directly connected to therapeutic effect., can by literature search and clinical trial registration data statistics
To find most of MSCs used in clinical research as autologous bone marrow source, and autologous bone marrow source MSCs manufacturing cycle
At least needed for 3 week, this just constrains applications of the MSCs in Patients With Acute Myocardial Infarction.In addition, in vivo residing for MSCs
Microenvironment is hypoxemia or anoxic, and especially in the tissue such as marrow, fat, oxygen concentration is extremely low (1-8%).It is and current
The condition of MSCs amplification in vitro cultures is oxygen-enriched (O2Concentration be 21%), microenvironment in Process of in vitro residing for MSCs with
There is larger difference in the tissue microenvironment in its source, so may result in MSCs cellularity, certain change occurs for function.
The MSCs cultivated under excess oxygen is infused into site of myocardial infarction, and this is a micro-environmental hypoxia, and the vigor of stem cell also can
Produce negative regulation effect.
The present invention relates to the preparation and application of hypoxic tolerance type umbilical cord derived mesenchymal stem cell, hypoxic tolerance of the invention
Type umbilical cord derived mesenchymal stem cell prepares easy, easy large-scale production, and can remain higher by freezing, after recovery processing
Stability, can avoid because autologous MSCs individuation difference causes the resultant error of preclinical study.Meanwhile with richness
The MSCs cultivated under the conditions of oxygen is compared, and hypoxic tolerance type MSCs has stronger paracrine function and survival ability, can be effective
Promote angiogenesis, and can survive for a long time in vivo.Therefore the hypoxic tolerance type umbilical cord derived mesenchymal stem cell of the present invention
The problems such as preferably resolving on cell manufacturing cycle, time-to-live, paracrine ability, improve stem cell stability and
Curative effect, open the frontier of stem cell preclinical study.
The content of the invention
Based on background above, it is an object of the invention to provide a kind of preparation method of hypoxic tolerance type mescenchymal stem cell
And its application.
The purpose of the present invention is achieved through the following technical solutions：
A kind of preparation method of hypoxic tolerance type mescenchymal stem cell, comprises the following steps：(1) umbilical cord is cut into 1-
1.5cm segment, umbilical vein and arteria umbilicalis are removed, strip magnificent Tong Shi glue；(2) the magnificent Tong Shi glue stripped is put into one to fill
Cleaned in PBS culture dish, then magnificent Tong Shi glue is cut into 1-3mm3The tissue block of size, it is inoculated with 0.5-1.0cm spacing
In culture dish；(3) room temperature adds 10mL complete mediums after placing 15min, is put into hypoxemia incubator and cultivates, CO2Concentration
Control as 5%, O2Concentration control is 1-6%, O2Concentration passes through N2Balance, temperature are 37 DEG C；(4) treat that the fusion rate of cell reaches
When more than 80%, cell is collected with Trypsin Induced, according to 5000cells/cm2Density is by passage to multiple blake bottles
In continue to cultivate；(5) after reaching P3-10 generations, with trypsin digestion and cell, 300g centrifugations 5min is collected, finally counted
And freeze.
Further, hypoxic tolerance type mescenchymal stem cell can be placed in Thermo Fisher HeracelTM150i incubators
In, CO2Concentration control is 5%, O2Concentration control is 1-6%, O2Concentration passes through N2Balance；And normal type mescenchymal stem cell is then
It is placed in the incubators of Thermo Forma 311, CO2Concentration control is 5%, O2Concentration control is 21%.
In embodiments of the invention 2-4, hypoxic tolerance type mescenchymal stem cell uses 1% oxygen concentration culture, normal type
The 4th generation (P4) cell is used with hypoxic tolerance type mescenchymal stem cell.
In embodiments of the invention 4, the solution medium of normal type and the use of hypoxic tolerance type mesenchymal stem cell transplantation
For PBS, cell density is 1 × 106cells/mL。
Present invention also offers application of the hypoxic tolerance type mescenchymal stem cell in acute myocardial infarction AMI is treated.
The creative discovery of the present inventor, compared with the prior art, there is the advantages of following prominent：
(1) hypoxic tolerance type mescenchymal stem cell of the invention is prepared simple, and has good stability.
(2) hypoxic tolerance type mescenchymal stem cell survival ability of the invention is strong, and apoptosis is not easy under anaerobic environment.
(3) hypoxic tolerance type mescenchymal stem cell of the present invention can discharge more compared with normal type mescenchymal stem cell
VEGF and HGF, and the angiogenesis at myocardial damage position can be effectively facilitated.
(4) hypoxic tolerance type mescenchymal stem cell of the invention promotes the ability of Myocardial Regeneration to be filled between being significantly higher than normal type
Matter stem cell, the present invention in hypoxic tolerance type mescenchymal stem cell can effectively treat acute myocardial infarction AMI.
Brief description of the drawings
The present invention is further detailed explanation with reference to the accompanying drawings and detailed description.
Fig. 1 is different oxygen concentrations for the multiplication capacity of mescenchymal stem cell and the testing result of paracrine ability；Its
Middle A is the proliferation times that mescenchymal stem cell is counted by cell count；B is that ELISA detects cell factor in culture supernatant
VEGF and HGF burst size, C are the expression quantity that western blotting detect HIF-1 α in mescenchymal stem cell；
Fig. 2 is the flow cytometer showed result figure of the surface marker of hypoxic tolerance type mescenchymal stem cell (it-MSC)；A is P4
For it-MSC flow cytometer showed result figure, B is flow cytometer showed result figures of the P10 for it-MSC；
Fig. 3 is normal type mescenchymal stem cell (N-MSC) and hypoxic tolerance type mescenchymal stem cell (it-MSC) serious
The flow cytometer detection result of cell death situation under anoxia condition；
Fig. 4 is the detection knot for promoting angiogenesis in N-MSC and it-MSC Stem Cell Transplantation in Treating Acute Myocardial Infarction rat models
Fruit；Wherein A is CD31, Hoechst immunofluorescence dyeing result of rat heart muscle tissue, and B is the CD31 dyeing in quantitative analysis A
Positive region；Vehicle represents solution medium control group；
Fig. 5 is the detection knot for promoting Myocardial Regeneration in N-MSC and it-MSC Stem Cell Transplantation in Treating Acute Myocardial Infarction rat models
Fruit；Wherein A is Ki67, TnI and Hoechst immunofluorescence dyeing result of rat heart muscle tissue, and B is in quantitative analysis A
Ki67 stained positives region.
Embodiment
A kind of non-limiting embodiment of hypoxic tolerance type mescenchymal stem cell prepared in accordance with the present invention is given below.
1 different O of embodiment2Influence of the concentration for mescenchymal stem cell multiplication capacity and paracrine function
Umbilical cord is cut into 1-1.5cm segment, then cut off with eye scissors along umbilical vein chamber, umbilical vein and arteria umbilicalis are gone
Remove, strip magnificent Tong Shi glue.The magnificent Tong Shi glue stripped is put into a new culture dish for filling PBS and cleaned, with another new eye
Section cuts is cut into 1-3mm by magnificent Tong Shi glue3The tissue block of size, it is inoculated in 0.5-1.0cm spacing in culture dish, room temperature is put
Put 15min and then add 10mL complete mediums.By above-mentioned tissue block be placed in different oxygen concentrations (21%, 16%, 11%,
6%th, 1%) cultivated in environment, digested and collected thin using trypsase TrypLE when the fusion rate of cell reaches more than 80%
Born of the same parents, then it is inoculated into respectively in 24 orifice plates again, it is each to be inoculated with 9 holes, inoculum density 4000cells/cm2；Oxygen concentration still maintains
The numerical value of prior-generation cell.
Different time points (24,48 and 72h) collect cell respectively, are carried out using Countstar full-automatic cells calculating instrument
Count, as a result as shown in Figure 1A, the growth rate of (6%, 1%) mescenchymal stem cell is slightly below under excess oxygen under hypoxia condition
Mescenchymal stem cell (21%, 16%, 11%), but this species diversity is not statistically significant.72h cells and supernatants are collected,
Use VEGF Human ELISA Kit (ThermoFisher, Cat#BMS277-2) and HGF Human ELISA Kit
(ThermoFisher, Cat#KAC2211) kit detects VEGF and HGF burst size, as a result as shown in Figure 1B, compared to oxygen-enriched
Under the conditions of mescenchymal stem cell (21%, 16%, 11%), the mescenchymal stem cell (6%, 1%) under hypoxia condition can discharge
More VEGF and HGF, this species diversity have statistical significance.Because VEGF and HGF can promote angiogenesis, prompt anoxic resistance to
There is stronger angiogenesis promoting ability by type mescenchymal stem cell.
Further, extract above-mentioned each cell protein and carry out western blotting detections, it is as a result as shown in Figure 1 C, low
Oxygen can raise the expression of transcription factor HIF-1 α in mescenchymal stem cell, and this is exactly that VEGF and HGF burst sizes are increased basic former
Cause.Based on above result of the test, oxygen concentration is (1-6%) used by hypoxic tolerance type mescenchymal stem cell of the invention,
In this oxygen concentration range, mescenchymal stem cell has stronger paracrine function, and its multiplication capacity does not substantially change.
The detection of the hypoxic tolerance type mescenchymal stem cell stability of embodiment 2
Hypoxic tolerance type mescenchymal stem cell (it-MSC) is put into Thermo Fisher HeracellTM150i hypoxemia cultures
Cultivated in case, CO2Concentration control is 5%, O2Concentration control is 1%, and temperature is 37 DEG C；Normal type mescenchymal stem cell (N-MSC)
It is put into the oxygen-enriched incubator cultures of Thermo Forma 311, CO2Concentration control is 5%, O2Concentration control is 21%, temperature 37
℃.When the fusion rate of cell reaches more than 80%, digested with trypsase TrypLE and collect cell, according to 5000cells/
cm2Passage is continued to cultivate by density into multiple T175 blake bottles, and condition of culture is the same as above-mentioned consistent.
After reaching generation in 3-10 generations (P3-P10), digested with trypsase TrypLE and collect cell, 300g centrifugation 5min, meter
Number and freeze-stored cell.The standard formulated according to international cell therapy association (ISCT), to above-mentioned 4th generation (P4) and the 10th generation
(P10) hypoxic tolerance type mescenchymal stem cell is identified, with the expression of Flow cytometry cell surface marker.
From Fig. 2A, B, in P4 generations and P10 prepared by the present embodiment, have for hypoxic tolerance type mescenchymal stem cell
Similar phenotype is both high to express CD73, CD90, CD105 (positive rate>95%), without expressing CD11b, CD14 and CD34
(positive rate<2%).Result above shows that the hypoxic tolerance type mescenchymal stem cell of the present embodiment has good stability, passes
Generation number has not significant impact to its phenotype.
Influence of the severe depletion of oxygen microenvironment of embodiment 3 for mescenchymal stem cell apoptosis
By P4 for normal type and hypoxic tolerance type mescenchymal stem cell, it is inoculated into respectively in 24 orifice plates, inoculum density is
4000cells/cm2；Then Thermo Fisher Heracell are placed inTMCultivated in 150i hypoxemia incubators, oxygen concentration
0.1% is adjusted to, the microenvironment of severe depletion of oxygen after myocardial infarction occurs with simulation.After cultivating 48h, cell is collected, is used
Annexin-V FITC Apoptosis Assay Kit detect normal type and the dead feelings of hypoxic tolerance type mescenchymal stem cell
Condition.
As a result as shown in figure 3, after meeting with the stimulation of severe depletion of oxygen obvious cell can occur for normal type mescenchymal stem cell
It is dead that (the mono- positive cell ratios of PI are that the mono- positive cell ratios of 13.0%, Annexin V are 0.6%, Annexin V and PI double
14.0%) positive cell ratio is, and hypoxic tolerance type mescenchymal stem cell can preferably adapt to the environment of severe depletion of oxygen, and cell is dead
Die ratio it is smaller (the mono- positive cell ratios of PI be the mono- positive cell ratios of 1.8%, Annexin V be 2.6%, Annexin V and
2.5%) PI double positive cells ratio is.Result above shows hypoxic tolerance type mescenchymal stem cell in the environment of severe depletion of oxygen
With stronger survival ability.
Therapeutic effect of the hypoxic tolerance type mescenchymal stem cell of embodiment 4 to acute myocardial infarction in rats
(1) structure of acute myocardial infarction of rat model
Male athymic rat (Sprague-Dawley Rats, SD rat) is bought, chooses the SD that body weight is 200-280g
Rat is tested, and ami model is built by arteria coroaria sinistra ligation operation.
SD rats are anaesthetized using the mode of intraperitoneal injection 30mg/kg yellow Jackets, and it is auxiliary with adjustable volume section lung ventilator
Help breathing.Then expose its heart by left chest throacotomy, will be between pulmonary artery and left ventricle with 7-0 prolene sutures
Arteria coroaria sinistra transience ligation, cause myocardial ischemic injury.
(2) transplantation treatment of mescenchymal stem cell
24 ami model rats are randomly divided into 3 groups, every group 8；Respectively solution medium (Vehicle)
Normal type mescenchymal stem cell treatment group (N-MSC) and hypoxic tolerance type mescenchymal stem cell treatment group (it-MSC).
Carry out intramyocardial injection within the 4th day after acute myocardial infarction model architecture：By rat anesthesia and chest is carried out out, is then made
With 30-gauge syringe needles by the solution medium or mescenchymal stem cell (1 × 10 in the 100 above-mentioned packets of μ L6Cells/mL the heart) is thrown into
It is intramuscular.
(3) mescenchymal stem cell treatment promotes the assessment of angiogenesis ability
Mescenchymal stem cell puts to death rat after treating 28 days, take out cardiac muscular tissue and freezing microtome section is made.Choose myocardial infarction
The section of fringe region, carry out CD31 immunofluorescence dyeings.Histotomy using 10% formalin fix 10min, 0.2%
Triton-X100 penetrating 15min, 3%BSA closing 1h, then add 4 DEG C of overnight incubations of CD31 antibody, are eventually adding fluorescence two
Anti- incubation at room temperature 1h.Nucleus is dyed using Hoechst 33258.Coloration result is using fluorescence microscope and claps
According to as a result as shown in Figure 4.It was found from Fig. 4 A and B, relative to Vehicle control groups, N-MSC treatment groups and it-MSC treatment groups
Middle rat myocardium from injury position vessel density substantially increases；And it-MSC promotes the ability of angiogenesis to be significantly higher than N-MSC.By
This is visible, and the hypoxic tolerance type mescenchymal stem cell in the present invention can effectively facilitate angiogenesis.
(4) mescenchymal stem cell treatment promotes the assessment of cardiac muscle mitochondria ability
Mescenchymal stem cell puts to death rat after treating 28 days, take out cardiac muscular tissue and freezing microtome section is made.Choose myocardial infarction
The section of fringe region, carry out troponin I (TnI) and Ki67 immunofluorescence dyeings.Histotomy uses 10% formalin
Fixed 10min, 0.2%Triton-X100 penetrating 15min, 3%BSA closing 1h, then add Anti-Cardiac
4 DEG C of overnight incubations of Troponin I antibody and Anti-Ki67 antibody, are eventually adding fluorescence secondary antibody incubation at room temperature 1h.Nucleus makes
Dyed with Hoechst 33258.
Coloration result is using fluorescence microscope and takes pictures, as a result as shown in Figure 5.It was found from Fig. 5 A and B, N-MSC is controlled
There is a small amount of newborn cardiac muscle cell in rat myocardium from injury position in treatment group and it-MSC treatment groups；And it-MSC promotes cardiac muscle
The ability of regeneration is significantly higher than N-MSC.As can be seen here, the hypoxic tolerance type mescenchymal stem cell in the present invention can effectively treat urgency
Property myocardial infarction.
The foregoing is only a preferred embodiment of the present invention, but protection scope of the present invention be not limited thereto,
Any one skilled in the art the invention discloses technical scope in, the change or replacement that can readily occur in,
It should all be included within the scope of the present invention.Therefore, protection scope of the present invention should be with the protection model of claims
Enclose and be defined.
Claims (4)
1. a kind of preparation method of hypoxic tolerance type mescenchymal stem cell, comprises the following steps：(1) umbilical cord is cut into 1-1.5cm
Segment, umbilical vein and arteria umbilicalis are removed, strip magnificent Tong Shi glue；(2) the magnificent Tong Shi glue stripped is put into one and fills PBS's
Cleaned in culture dish, then magnificent Tong Shi glue is cut into 1-3mm3The tissue block of size, culture is inoculated in 0.5-1.0cm spacing
In ware；(3) room temperature adds 10mL complete mediums after placing 15min, is put into hypoxemia incubator and cultivates, CO2Concentration controls
5%, O2Concentration control is 1-6%, O2Concentration passes through N2Balance, temperature are 37 DEG C；(4) treat cell fusion rate reach 80% with
When upper, cell was collected with Trypsin Induced, according to 5000cells/cm2Density continues passage into multiple blake bottles
Culture；(5) after reaching 3-10 generations (P3-P10), with trypsin digestion and cell, 300g centrifugations 5min is collected, finally counted
Count and freeze.
A kind of 2. preparation method of hypoxic tolerance type mescenchymal stem cell according to claim 1, it is preferred that O2Concentration control
It is made as 1%.
3. the preparation method of a kind of hypoxic tolerance type mescenchymal stem cell according to claim 1, it is preferred that mesenchyma is done
After passage reaches forth generation (P4), with trypsin digestion and cell, 300g centrifugations 5min is collected, finally counts and freeze
Deposit.
4. a kind of hypoxic tolerance type mescenchymal stem cell as described in claims 1 to 3 is any moves in treatment acute myocardial infarction AMI
Application in thing model.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710704337.4A CN107384858A (en) | 2017-08-17 | 2017-08-17 | A kind of preparation method and applications of hypoxic tolerance type mescenchymal stem cell |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710704337.4A CN107384858A (en) | 2017-08-17 | 2017-08-17 | A kind of preparation method and applications of hypoxic tolerance type mescenchymal stem cell |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107384858A true CN107384858A (en) | 2017-11-24 |
Family
ID=60353026
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710704337.4A Pending CN107384858A (en) | 2017-08-17 | 2017-08-17 | A kind of preparation method and applications of hypoxic tolerance type mescenchymal stem cell |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107384858A (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108251359A (en) * | 2017-12-20 | 2018-07-06 | 上海华新生物高技术有限公司 | A kind of mesenchymal stem cell serum-free culture medium and cultural method |
| CN110079496A (en) * | 2018-01-25 | 2019-08-02 | 左凤琼 | A kind of method of hypoxemia culture menses source Endometrial stem cell |
| CN112430626A (en) * | 2020-11-27 | 2021-03-02 | 成都康景生物科技有限公司 | Genetically modified umbilical mesenchymal stem cell, preparation method and application |
| CN114276990A (en) * | 2022-01-13 | 2022-04-05 | 协和华东干细胞基因工程有限公司 | Method and system for improving mesenchymal stem cell repair capacity |
| CN114480268A (en) * | 2022-01-21 | 2022-05-13 | 深圳市茵冠生物科技有限公司 | Preparation method of human umbilical cord mesenchymal stem cells |
| CN114940975A (en) * | 2022-06-20 | 2022-08-26 | 中国人民解放军总医院 | TED-deleted cell line and application thereof |
| CN116083352A (en) * | 2023-03-26 | 2023-05-09 | 广州冠邦生物科技有限公司 | Method for in-vitro directional differentiation of mesenchymal stem cells into myocardial cells |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104781394A (en) * | 2012-09-03 | 2015-07-15 | Medipost株式会社 | Method for culturing mesenchymal stem cells |
| CN104946586A (en) * | 2015-07-20 | 2015-09-30 | 龚伟 | Pretreatment method of mesenchymal stem cells and preparation obtained from mesenchymal stem cells |
| CN106244532A (en) * | 2016-09-08 | 2016-12-21 | 石家庄融和生物科技有限责任公司 | The preparation method of people source umbilical cord mesenchymal stem cells |
-
2017
- 2017-08-17 CN CN201710704337.4A patent/CN107384858A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104781394A (en) * | 2012-09-03 | 2015-07-15 | Medipost株式会社 | Method for culturing mesenchymal stem cells |
| CN104946586A (en) * | 2015-07-20 | 2015-09-30 | 龚伟 | Pretreatment method of mesenchymal stem cells and preparation obtained from mesenchymal stem cells |
| CN106244532A (en) * | 2016-09-08 | 2016-12-21 | 石家庄融和生物科技有限责任公司 | The preparation method of people source umbilical cord mesenchymal stem cells |
Non-Patent Citations (2)
| Title |
|---|
| USHA NEKANTI等: "Increased proliferation and analysis of differential gene espression in human wharton"s jelly-derived mesenchymal stromal cells under hypoxia", 《INT J BIOL SCI》 * |
| XINYANG HU等: "Transplantation of hypoxia-preconditioned mesenchymal stem cells improves infarcted heart function via enhanced survival of implanted cells and angiogenesis", 《THE JOURNAL OF THORACIC AND CARDIOVASCULAR SURGERY》 * |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108251359A (en) * | 2017-12-20 | 2018-07-06 | 上海华新生物高技术有限公司 | A kind of mesenchymal stem cell serum-free culture medium and cultural method |
| CN108251359B (en) * | 2017-12-20 | 2021-07-09 | 上海华新生物高技术有限公司 | Mesenchymal stem cell serum-free medium and culture method |
| CN110079496A (en) * | 2018-01-25 | 2019-08-02 | 左凤琼 | A kind of method of hypoxemia culture menses source Endometrial stem cell |
| CN112430626A (en) * | 2020-11-27 | 2021-03-02 | 成都康景生物科技有限公司 | Genetically modified umbilical mesenchymal stem cell, preparation method and application |
| CN114276990A (en) * | 2022-01-13 | 2022-04-05 | 协和华东干细胞基因工程有限公司 | Method and system for improving mesenchymal stem cell repair capacity |
| CN114480268A (en) * | 2022-01-21 | 2022-05-13 | 深圳市茵冠生物科技有限公司 | Preparation method of human umbilical cord mesenchymal stem cells |
| CN114480268B (en) * | 2022-01-21 | 2024-01-30 | 深圳市茵冠生物科技有限公司 | Preparation method of human umbilical cord mesenchymal stem cells |
| CN114940975A (en) * | 2022-06-20 | 2022-08-26 | 中国人民解放军总医院 | TED-deleted cell line and application thereof |
| CN116083352A (en) * | 2023-03-26 | 2023-05-09 | 广州冠邦生物科技有限公司 | Method for in-vitro directional differentiation of mesenchymal stem cells into myocardial cells |
| CN116083352B (en) * | 2023-03-26 | 2023-11-14 | 河北意和医学检验实验室有限公司 | Method for in-vitro directional differentiation of mesenchymal stem cells into myocardial cells |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107384858A (en) | A kind of preparation method and applications of hypoxic tolerance type mescenchymal stem cell | |
| CN106109496B (en) | Human umbilical cord mesenchymal stem cells extract freeze-drying powder and preparation method | |
| CN106982821A (en) | Umbilical cord mesenchymal stem cells clinic freezes protection liquid composition and application thereof | |
| CN104195098A (en) | Dendrobium officinale stem cell and isolated culture method thereof | |
| CN104958319A (en) | Mesenchymal stem cell and cytokine preparation having treatment effects on premature ovarian failures and perimenopausal syndromes, and preparing method for preparation | |
| CN110564682B (en) | Method for large-scale production of human adipose-derived mesenchymal stem cell exosomes | |
| CN107028980A (en) | Pharmaceutical composition for treating heart disease | |
| CN105687244B (en) | A kind of preparation, preparation method and its application | |
| CN110157666A (en) | Umbilical cord mesenchymal stem cells MSCs and its cultural method and application | |
| CN106237313A (en) | A kind of umbilical cord mesenchymal stem cells injection and its preparation method and application | |
| CN105820998A (en) | Isolation extraction and culture method for human adipose-derived stem cells (ADSCs) for clinical back-transfusion grade cell therapy | |
| CN106511387A (en) | Preparing method and application of chicken embryo extracts | |
| CN109464463A (en) | The characteristic of the anti-skin epidermal cells oxidation of mescenchymal stem cell excretion body and its preparation method of preparation | |
| CN106539821A (en) | A kind of stem cell excretion body preparation and its preparation method and application | |
| CN105395571A (en) | New application of small exosome secreted by mesenchymal stem cells in treatment of myocardial infarction | |
| CN106566803A (en) | Culture solution, application of culture solution and method for culturing umbilical cord mesenchymal stem cells | |
| CN109646457A (en) | A kind of application of umbilical cord mesenchymal stem cells in preparation treatment acute lung injury drug | |
| CN107267452A (en) | A kind of method for resuscitation of dental pulp stem cell resuscitation fluid and dental pulp stem cell | |
| CN107488627A (en) | A kind of biological gel for treating intractable skin injury and its application | |
| CN108715831A (en) | Application of the mescenchymal stem cell conditioned medium in the cellular damage for improving cardiomyocytes induced by hypoxia and reoxygenation | |
| CN106606512B (en) | Mixed cell preparation for treating myocardial infarction and preparation method and application thereof | |
| CN108865985A (en) | A kind of method of the pre- epithelial-mesenchymal conversion of stem cell source excretion soma | |
| CN106491652A (en) | Goose eggembryosin preparation method and application | |
| CN109554454A (en) | A method of freeze-dried powder hair regrowth is evaluated by measurement cytokine content | |
| Li et al. | Effective component of Salvia miltiorrhiza in promoting cardiomyogenic differentiation of human placenta‑derived mesenchymal stem cells |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20171124 |