CN109402048A - The method for building up of " humanized " arrhythmogenic right ventricular cardiomyopathy disease model - Google Patents
The method for building up of " humanized " arrhythmogenic right ventricular cardiomyopathy disease model Download PDFInfo
- Publication number
- CN109402048A CN109402048A CN201811187690.0A CN201811187690A CN109402048A CN 109402048 A CN109402048 A CN 109402048A CN 201811187690 A CN201811187690 A CN 201811187690A CN 109402048 A CN109402048 A CN 109402048A
- Authority
- CN
- China
- Prior art keywords
- culture medium
- cell
- arvc
- day
- humanized
- 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
- C12N5/0657—Cardiomyocytes; Heart 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
- C12N2500/90—Serum-free medium, which may still contain naturally-sourced components
-
- 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
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/30—Hormones
- C12N2501/33—Insulin
-
- 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
- C12N2506/00—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells
- C12N2506/45—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells from artificially induced pluripotent 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
- C12N2533/00—Supports or coatings for cell culture, characterised by material
- C12N2533/50—Proteins
Abstract
The invention discloses a kind of method for building up of " humanized " arrhythmogenic right ventricular cardiomyopathy (ARVC) disease model.This method is that ARVC patient skin is carried out originally culture to obtain fibroblast, acquisition ARVC patient-specific is induced to induce multi-potent stem cell (iPSC) by nonconformity sendai virus, the cardiac muscle cell for being divided into ARVC patient-specific is directed it, again to establish stable external cardiac muscle cell's disease model.The present invention can establish stable humanized's ARVC model based on iPSC technology, and form and function meets ARVC pathological characters, which is suitable for ARVC related pathologies Mechanism Study and individuation drug screening.
Description
Technical field
The invention belongs to field of biomedicine technology, disclose a kind of " humanized " arrhythmogenic right ventricular cardiomyopathy
(ARVC) method for building up of disease model can be used for the related pathologies Mechanism Study of ARVC and its treat and prevent grinding for drug
Hair.
Background technique
Arrhythmogenic right ventricular cardiomyopathy (Arrhythmogenic right ventricular
Cardiomyopathy, ARVC) it is a kind of cardiomyopathy, male adults are more common in, male to female ratio is about 2.7:1.The disease pathology
Feature is myocardial atrophy and fibrofatty tissue's substitution in right ventricle, clinical often to show as right ventricle expansion, arrhythmia cordis, sudden
Dead and heart failure, is the common cause of the death in Exercise-related sudden death.30%~50% ARVC case has familial, main to show
For autosomal dominant inheritance, genepenetrance is different.It is currently known at least nine ARVC related gene, wherein most for coding desmosome
The gene of albumen.So far, it is not known about the pathogenetic Study on Molecular Mechanism of ARVC disease, and there is very big dispute, it is main
It to be the research based on mouse disease model.However, mouse and the mankind there are great differences on various physiological systems (such as people
The Beating Rate of cardiac muscle cell is 60 beats/min, and the Beating Rate of mouse is 400-600 beats/min), many results of study cannot
Directly apply to human diseases.Therefore, " humanized " cardiac muscle cell model how is established for cardiovascular diseases such as ARVC
Research it is particularly significant.
People induces multi-potent stem cell (human induced Pluripotent Stem Cells, hiPSCs)
Translational medicine and regenerative medicine provide powerful power and revolutionary Mode change, and stem cell of having furthered is used for clinical disease
The distance for the treatment of.HiPSCs is widely used in the research of cardiovascular disease, is ground in disease model foundation, disease molecules mechanism
Study carefully and new drug development etc. has huge potential value.Compared with people's adult stem cell, one of hiPSCs is obvious excellent
Gesture is its versatility, and exactly this versatility, and imparting it can be divided into as the energy of people's any one cell in vivo
Power, such as cardiac muscle cell, vascular cell, nerve cell, liver cell.For Cardiovascular Disease Study, the primary cardiac muscle of people
Cell is optimal cellular resources, however is difficult to obtain, and condition of in vitro culture is harsh, therefore its application is by the very day of one's doom
System.Using myocardium directed differentiation technology, hiPSCs can be broken up becomes cardiac muscle cell, cardiac muscle cell derived from these hiPSC
(hiPSC-CMs) human genome is carried, there is environment in " humanized " physiology, obtain relatively easy and can train for a long time in vitro
Support, be currently known with the most similar cellular resources of people's primary cardiomyocytes.
Summary of the invention
The purpose of the present invention is overcome the deficiencies of the prior art and provide " humanized " arrhythmogenic right ventricular cardiomyopathy
The method for building up of disease (ARVC) disease model.
The method for building up of " humanized " arrhythmogenic right ventricular cardiomyopathy disease model includes the following steps:
1) originally culture obtains fibroblast
By the PBS+1% mycillin rinse of ARVC patient skin tissue piece, PBS is siphoned away, clostridiopetidase A is added in 37 DEG C
Digestion 6 hours.DMEM+10%FBS+1% mycillin culture medium is then added, is placed in incubator culture, waits skin histology
It is adherent, liquid was changed every 4 days.When in culture dish fibroblast cover with after obtain P0For fibroblast, freezes conservation and pass on
Amplification cultivation.
Wherein the PBS is phosphate balance physiological saline, and DMEM is well known in the art a kind of containing various amino acid
With the culture medium of glucose, the percentage in PBS+1% mycillin and DMEM+10%FBS+1% mycillin culture medium is equal
It calculates by volume.
2) foundation of ARVC patient-specific iPS cell strain
Fibroblast kind is entered in orifice plate, the cell for selecting 1 hole just to cover with carries out sendai virus infection, after 24 hours
It changes DMEM+10%FBS+1% mycillin culture medium into, liquid was changed every 1 day, cell was passed in the 7th day and is covered in advance
In the culture dish of Matrigel, while culture medium is changed to mTeSR.
By single clone's picking to in the orifice plate for being covered with Matrigel in advance, cell is ARVC-iPSCP0 generation, choosing at this time
It selects the clone that cell is fine and close, periphery does not break up to pass on into the orifice plate for being covered with Matrigel in advance, cell is P1 generation at this time；Choosing
Cell is fine and close, periphery does not break up 3 clones passage is selected to in the orifice plate for being covered with Matrigel in advance, cell is P2 at this time
Cell is passaged to P20 generation always in the orifice plate and every 1 generation all carries out conservation by generation.
Wherein Matrigel is basement membrane matrix glue, and mTeSR is human pluripotent stem cells serum free medium；
3) acquisition of ARVC patient-specific cardiac muscle cell
2D single layer using small molecule compound induction breaks up scheme, basic fundamental route are as follows: the 1st day completion iPSCs
Break up to mesoblastic induction, the 1-3 days completion mesoderms are to the mesoblastic induction transdifferentiation of heart, embryo in completion in the 4-7 days
The induction differentiation of layer cells into cardiomyocytes is mainly the maturation of cardiac muscle cell after the 7th day.IPSCs is pressed into 1:8-1 first:
12 ratios are seeded in 6 orifice plates, replace culture medium daily, can carry out differentiation operation when cell density reaches 80% or so, are being divided
Change the same day, exhaust outmoded culture medium, Myocardium Differentiation culture medium (RPMI+B27-Insulin) cleans one time, and every hole, which is added, contains 8 μM
CHIR differential medium 2ml；Myocardium Differentiation culture medium is supplemented after 1 day；Myocardium Differentiation culture medium, every hole are replaced with after 2 days
2ml；3rd day, the Myocardium Differentiation culture medium 2ml of the IWR containing 5 μM was added in every hole, continued culture 2 days, the 4th day supplement Myocardium Differentiation
Culture medium；It is replaced with Myocardium Differentiation culture medium within 5th day；Myocardial cells culture base (RPMI+B27+ is replaced with after 7th day
Insulin), at this time if the cardiac muscle cell to beat in flakes well can be seen in differentiation effect, show " humanized " proarrhythmia
Property arrhythmogenic disease model is successfully established.
Preferably, in the RPMI+B27-Insulin culture medium, the 2% of the total culture solution total amount of B27-Insulin Zhan.
Preferably, in the RPMI+B27+Insulin culture medium, the 2% of the total culture solution total amount of B27+Insulin Zhan.
Preferably, the magnitude of recruitment of supplement RPMI+B27-Insulin culture medium is on original culture medium in the step 3)
The half of former culture medium.
The present invention can establish stable humanized's ARVC disease model based on iPSC technology, and form and function meets ARVC
Feature, the model are suitable for ARVC related pathologies Mechanism Study and individuation drug screening.
Detailed description of the invention
Fig. 1: iPSC versatility qualification result；
Fig. 2: the myofilament texture colored graph of normal person's group and patient ARVC group cardiac muscle cell；
The electro physiology phenotype of Fig. 3: ARVC patient group cardiac muscle cell；
The fat deposition phenotype of Fig. 4: ARVC patient group cardiac muscle cell.
Specific embodiment
1) separation and its originally culture of skin fibroblasts: after obtaining skin, adipose tissue is cut off and skin group
It knits and is cut into small pieces, with PBS+1% mycillin rinse 1 time, addition clostridiopetidase A digests 6 hours at 37 DEG C after siphoning away PBS, digestion knot
DMEM+10%FBS+1% mycillin culture medium is added in Shu Hou, is put into incubator culture, waits skin group adherent, every 4 days
Change liquid.It is sure not to blow skin histology, it can be observed that fibroblast after 1 week.
2) fibroblast the foundation of ARVC patient-specific iPS cell strain: is pressed 1 × 104,2×104,3×104,4×
104, 6 × 104,8×104Gradient be inoculated in 48 orifice plates, the 2nd day observation after select 1 hole just to cover with cell carry out celestial platform
Virus infection, changes DMEM+10%FBS+1% mycillin culture medium into after 24 hours, the next day change liquid, cell was passed in the 7th day
With in the 60-mm culture dish for being covered with Matrigel in advance, while culture medium uses mTeSR (STEMCELL Technologies) instead.
Observation cell daily, after cloning suitable picking, will single clone's picking to in 96 orifice plates for being covered with Matrigel in advance, this
When cell be P0 generation.The undesirable cell of cell state is abandoned, the excellent clone's passage of selection state to being covered in advance
In 24 orifice plates of Matrigel, cell is P1 generation at this time.3 clones in the best state are selected to pass on to being covered in advance
In 12 orifice plates of Matrigel, cell is P2 generation at this time, and cell is passaged to P20 generation and every 1 Dai Doujin always in 12 orifice plates
Row conservation.Pass through immunofluorescence experiment NANOG, SSEA4, SOX2 and the Oct3/4 positive and alkaline phosphatase staining (AP
Staining) versatility (Fig. 1) of iPSC has been established in experimental identification.
3) acquisition of ARVC patient-specific cardiac muscle cell: the 2D single layer using small molecule compound induction breaks up scheme,
Its basic fundamental route are as follows: completion iPSCs broke up to mesoblastic induction in the 1st day, the 1-3 days completion mesoderm embryos into heart
The induction transdifferentiation of layer completes the induction differentiation of mesoderm cells into cardiomyocytes for the 4-7 days, is mainly cardiac muscle cell after the 7th day
Maturation.IPSCs is seeded in 6 orifice plates in 1:8-1:12 ratio first, replaces culture medium daily, when cell density reaches
Differentiation operation can be carried out to 80% or so, on the day of differentiation, exhausts outmoded culture medium, Myocardium Differentiation culture medium (RPMI+B27-
Insulin) (Gibco) is cleaned one time, and the differential medium 2ml of the CHIR (AXON) containing 8 μM is added in every hole；The heart is supplemented after 1 day
Flesh differential medium；Myocardium Differentiation culture medium, every hole 2ml are replaced with after 2 days；3rd day, the IWR containing 5 μM was added in every hole
(MERCK) Myocardium Differentiation culture medium 2ml continues culture 2 days, the 4th day supplement Myocardium Differentiation culture medium；Replacement in 5th day is deliberately
Flesh differential medium；Myocardial cells culture base (RPMI+B27+Insulin) (Gibco) is replaced with after 7th day, at this time if
The cardiac muscle cell to beat in flakes well can be seen in differentiation effect.
4) phenotypic evaluation of ARVC model: Cardiac-specific marker TNNT2 (Abcam) and α-actinin is applied
(Abcam) cardiac muscle cell's immunostaining is carried out, identifies that the cell that induction obtains is cardiac muscle cell (Fig. 2).Pass through patch clamp technique
The action potential that normal person's group and patient ARVC on individual cell level organize cardiac muscle cell is recorded, patient ARVC can be observed and organize cardiac muscle
There is apparent arrhythmia cordis phenotype (Fig. 3) in cell.It is generated by fat inducing factor inducing cardiomyocytes fat, passes through transmission
Electronic Speculum shoots intracellular fat deposition situation, and discovery patient ARVC compared with normal person organizes cardiac muscle cell organizes cell and dramatically increases
Fat drips exist, have apparent fat deposition phenotype (Fig. 4).The clinical phenotypes kissing that the above cell phenotype and patient occur
It closes.
5) application of ARVC model: result of study of the present invention shows that reliable, stable people can be established using iPSC technology
Source property ARVC disease model, form and function meet ARVC feature, which is suitable for the base of ARVC related pathologies mechanism
Plinth research, and patient can be reversed to organize in cardiac muscle cell arrhythmia cordis phenotype and fat deposition phenotype as index screening ARVC
The drug of disease treatment and prevention.
Claims (4)
1. a kind of method for building up of " humanized " arrhythmogenic right ventricular cardiomyopathy disease model, includes the following steps:
1) originally culture obtains fibroblast
By the PBS+1% mycillin rinse of ARVC patient skin tissue block, PBS is siphoned away, it is small in 37 DEG C of digestion 6 that clostridiopetidase A is added
When；DMEM+10%FBS+1% mycillin culture medium is then added, is placed in incubator culture, waits skin histology adherent, often
Changed liquid every 4 days, when in culture dish fibroblast cover with after obtain P0For fibroblast, freezes conservation and pass on amplification training
It supports；
2) foundation of ARVC patient-specific iPS cell strain
Fibroblast kind is entered in orifice plate, the cell for selecting 1 hole just to cover with carries out sendai virus infection, changes into after 24 hours
DMEM+10%FBS+1% mycillin culture medium changed liquid every 1 day, is passed to cell within the 7th day and is covered with Matrigel's in advance
In culture dish, while culture medium is changed to mTeSR；
By single clone's picking to in the orifice plate for being covered with Matrigel in advance, cell is ARVC-iPSCP0 generation at this time, and selection is thin
The clone that born of the same parents are fine and close, periphery does not break up is passed on into the orifice plate for being covered with Matrigel in advance, and cell is P1 generation at this time；Selection is thin
3 clones passage that born of the same parents are fine and close, periphery does not break up is to in the orifice plate for being covered with Matrigel in advance, and cell is P2 generation at this time,
Cell is passaged to P20 generation always in the orifice plate and every 1 generation all carries out conservation；
3) acquisition of ARVC patient-specific cardiac muscle cell
2D single layer using small molecule compound induction breaks up scheme, technology path are as follows: the 1st day completion iPSCs is to mesoderm
Induction differentiation, the 1-3 days completion mesoderms are to the mesoblastic induction transdifferentiation of heart, the 4-7 days completion mesoderm Cardiomyocytes
The induction of cell is broken up, and is the maturation of cardiac muscle cell after the 7th day；
Step 3) the concrete operations are as follows: be first seeded to iPSCs in 6 orifice plates in 1:8-1:12 ratio, daily replacement training
Base is supported, differentiation operation is carried out when cell density reaches 80%, on the day of differentiation, exhausts outmoded culture medium, Myocardium Differentiation culture medium
The differential medium 2ml of the CHIR containing 8 μM is added in cleaning one time, every hole；Myocardium Differentiation culture medium is supplemented after 1 day；It is replaced after 2 days
At myocardium differential medium, every hole 2ml；3rd day, the Myocardium Differentiation culture medium 2ml of the IWR containing 5 μM was added in every hole, continued to cultivate
2 days, the 4th day supplement Myocardium Differentiation culture medium；It is replaced with Myocardium Differentiation culture medium within 5th day；Cardioblast is replaced after 7th day
Culture medium, differentiation effect is well i.e. it can be seen that the cardiac muscle cell to beat in flakes, that is, show " humanized " arrhythmogenic at this time
Arrhythmogenic disease model, which is established, to be completed.
2. the method for building up of " humanized " arrhythmogenic right ventricular cardiomyopathy disease model according to claim 1,
It is characterized in that the Myocardium Differentiation culture medium is RPMI+B27-Insulin, wherein the total culture medium of B27minus Insulin Zhan
The 2% of total amount.
3. the method for building up of " humanized " arrhythmogenic right ventricular cardiomyopathy disease model according to claim 1,
It is characterized in that the myocardial cells culture base is RPMI+B27+Insulin, wherein the total culture medium total amount of B27+Insulin Zhan
2%.
4. the method for building up of " humanized " arrhythmogenic right ventricular cardiomyopathy disease model according to claim 1,
It is characterized in that the magnitude of recruitment for supplementing Myocardium Differentiation culture medium in the step 3) on original culture medium is the half of former culture medium.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811187690.0A CN109402048A (en) | 2018-10-11 | 2018-10-11 | The method for building up of " humanized " arrhythmogenic right ventricular cardiomyopathy disease model |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811187690.0A CN109402048A (en) | 2018-10-11 | 2018-10-11 | The method for building up of " humanized " arrhythmogenic right ventricular cardiomyopathy disease model |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109402048A true CN109402048A (en) | 2019-03-01 |
Family
ID=65467049
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811187690.0A Pending CN109402048A (en) | 2018-10-11 | 2018-10-11 | The method for building up of " humanized " arrhythmogenic right ventricular cardiomyopathy disease model |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109402048A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108913655A (en) * | 2018-07-16 | 2018-11-30 | 浙江大学 | The method for establishing " humanized " myocardial hypertrophy model based on multipotential stem cell technology |
| CN111575227A (en) * | 2020-04-08 | 2020-08-25 | 浙江大学 | Method for establishing human-derived diabetic cardiomyopathy model |
| CN113025654A (en) * | 2021-01-13 | 2021-06-25 | 河北医科大学 | Construction method of human induced pluripotent stem cell bank |
| CN115369077A (en) * | 2022-07-29 | 2022-11-22 | 佛山市中科律动生物科技有限公司 | MEFLC cell strain, and construction method and application thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102947444A (en) * | 2010-06-23 | 2013-02-27 | 帷幄生物技术公司 | Compositions and methods for re-programming cells without genetic modification for treatment of cardiovascular diseases |
| CN103987854A (en) * | 2011-07-21 | 2014-08-13 | 小利兰·斯坦福大学托管委员会 | Cardiomyocytes from induced pluripotent stem cells from patients and methods of use |
| CN107779429A (en) * | 2017-12-07 | 2018-03-09 | 陕西九州生物医药科技集团有限公司 | A kind of tissue-derived fibroblast quick separating cultural method of application on human skin |
| CN108060125A (en) * | 2018-02-07 | 2018-05-22 | 苏州大学 | Promote the method that pluripotent stem cell differentiation is cardiac muscle cell's maturation |
| CN108359636A (en) * | 2018-02-07 | 2018-08-03 | 苏州大学 | It is a kind of to improve the abductive approach that multipotential stem cell directed differentiation is cardiac muscle cell |
| US20200248145A1 (en) * | 2019-02-01 | 2020-08-06 | The University Of Hong Kong | Identification of subpopulations of cardiomyocytes |
-
2018
- 2018-10-11 CN CN201811187690.0A patent/CN109402048A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102947444A (en) * | 2010-06-23 | 2013-02-27 | 帷幄生物技术公司 | Compositions and methods for re-programming cells without genetic modification for treatment of cardiovascular diseases |
| CN103987854A (en) * | 2011-07-21 | 2014-08-13 | 小利兰·斯坦福大学托管委员会 | Cardiomyocytes from induced pluripotent stem cells from patients and methods of use |
| CN107779429A (en) * | 2017-12-07 | 2018-03-09 | 陕西九州生物医药科技集团有限公司 | A kind of tissue-derived fibroblast quick separating cultural method of application on human skin |
| CN108060125A (en) * | 2018-02-07 | 2018-05-22 | 苏州大学 | Promote the method that pluripotent stem cell differentiation is cardiac muscle cell's maturation |
| CN108359636A (en) * | 2018-02-07 | 2018-08-03 | 苏州大学 | It is a kind of to improve the abductive approach that multipotential stem cell directed differentiation is cardiac muscle cell |
| US20200248145A1 (en) * | 2019-02-01 | 2020-08-06 | The University Of Hong Kong | Identification of subpopulations of cardiomyocytes |
Non-Patent Citations (3)
| Title |
|---|
| FATIMA,A等: ""Derivation of induced pluripotent stem (iPS) cells from a patient with an arrhythmogenic right ventricular cardiomyopathy (ARVC)."", 《JOURNAL OF STEM CELLS & REGENERATIVE MEDICINE》 * |
| VERONICA SANCHEZ-FREIRE等: ""Effect of Human Donor Cell Source on Differentiation and Function of Cardiac Induced Pluripotent Stem Cells"", 《JOURNAL OF THE AMERICAN COLLEGE OF CARDIOLOGY》 * |
| 蒋彬等: ""致心律失常性右室心肌病患者的诱导性多能干细胞系的建立"", 《中国应用生理学杂志》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108913655A (en) * | 2018-07-16 | 2018-11-30 | 浙江大学 | The method for establishing " humanized " myocardial hypertrophy model based on multipotential stem cell technology |
| CN108913655B (en) * | 2018-07-16 | 2022-07-15 | 浙江大学 | Method for establishing human-derived myocardial hypertrophy model based on pluripotent stem cell technology |
| CN111575227A (en) * | 2020-04-08 | 2020-08-25 | 浙江大学 | Method for establishing human-derived diabetic cardiomyopathy model |
| CN113025654A (en) * | 2021-01-13 | 2021-06-25 | 河北医科大学 | Construction method of human induced pluripotent stem cell bank |
| CN115369077A (en) * | 2022-07-29 | 2022-11-22 | 佛山市中科律动生物科技有限公司 | MEFLC cell strain, and construction method and application thereof |
| CN115369077B (en) * | 2022-07-29 | 2023-07-18 | 佛山市中科律动生物科技有限公司 | MEFLC cell strain, construction method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Xue et al. | Functional integration of electrically active cardiac derivatives from genetically engineered human embryonic stem cells with quiescent recipient ventricular cardiomyocytes: insights into the development of cell-based pacemakers | |
| CN109402048A (en) | The method for building up of " humanized " arrhythmogenic right ventricular cardiomyopathy disease model | |
| Hansen et al. | Development of a drug screening platform based on engineered heart tissue | |
| Leri et al. | Role of cardiac stem cells in cardiac pathophysiology: a paradigm shift in human myocardial biology | |
| Lu et al. | Repopulation of decellularized mouse heart with human induced pluripotent stem cell-derived cardiovascular progenitor cells | |
| Caspi et al. | Transplantation of human embryonic stem cell-derived cardiomyocytes improves myocardial performance in infarcted rat hearts | |
| Planat-Benard et al. | Spontaneous cardiomyocyte differentiation from adipose tissue stroma cells | |
| Zhang et al. | Derivation and high engraftment of patient-specific cardiomyocyte sheet using induced pluripotent stem cells generated from adult cardiac fibroblast | |
| JPWO2014141528A1 (en) | Heart or vascular tissue type spheroid | |
| Blazeski et al. | Engineered heart slice model of arrhythmogenic cardiomyopathy using plakophilin-2 mutant myocytes | |
| CN109689858A (en) | Method for generating mesoderm and/or endothelium colony forming cell like cell with body vessel Forming ability | |
| CN104940997A (en) | Human tissue-engineered cardiac muscle tissue | |
| CN102161980B (en) | Method for culturing induced pluripotent stem cells by using human mesenchymal stem cells as trophoblast | |
| US9969978B2 (en) | Method for producing cardiomyocytes from human or mouse embryonic stem cells in a medium consisting of a serum-free medium and N2 supplement | |
| CN102719394B (en) | Method for constructing goat dermal fibroblast (DFB) line | |
| Rubach et al. | Mesenchymal Stem Cells and Their Conditioned Medium Improve Integration of Purified Induced Pluripotent Stem Cell–Derived Cardiomyocyte Clusters into Myocardial Tissue | |
| CN104491931B (en) | Sebaceous gland-containing skin tissue as well as formation method and application thereof | |
| CN103589686A (en) | Method of utilizing human umbilical cord mesenchymal stem cells as feed layer to culture human induced pluripotent stem cells | |
| Ma et al. | Location, isolation, and identification of mesenchymal stem cells from adult human sweat glands | |
| CN109963939A (en) | The derivative of pluripotent cell and self-renewing and application thereof | |
| Xi et al. | Fibroblasts support functional integration of purified embryonic stem cell-derived cardiomyocytes into avital myocardial tissue | |
| JP2016523514A (en) | Endocardial adult stem cells and method for producing the same | |
| McDonald et al. | Could cells from your nose fix your heart? Transplantation of olfactory stem cells in a rat model of cardiac infarction | |
| BR112020004517A2 (en) | stem cells derived from newborn pig and process for their preparation | |
| TW201504435A (en) | Quality control method for hair-follicle forming composition |
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 |
Application publication date: 20190301 |
|
| RJ01 | Rejection of invention patent application after publication |