CN110117570A - A kind of fibroblastic primary culture method of rheumatoid arthritis synovial - Google Patents
A kind of fibroblastic primary culture method of rheumatoid arthritis synovial Download PDFInfo
- Publication number
- CN110117570A CN110117570A CN201910458872.5A CN201910458872A CN110117570A CN 110117570 A CN110117570 A CN 110117570A CN 201910458872 A CN201910458872 A CN 201910458872A CN 110117570 A CN110117570 A CN 110117570A
- Authority
- CN
- China
- Prior art keywords
- synovial
- cell
- primary culture
- rheumatoid arthritis
- culture method
- 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.)
- Granted
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
- C12N5/0656—Adult fibroblasts
-
- 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
- C12N2509/00—Methods for the dissociation of cells, e.g. specific use of enzymes
Landscapes
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biomedical Technology (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Rheumatology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Cell Biology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention provides a kind of fibroblastic primary culture method of rheumatoid arthritis synovial, which comprises synovial tissue after pretreatment is sufficiently broken；III Collagenase Type and II Collagenase Type are added thereto and is blown and beaten；Individual cells are dispersed as after culture digestion；Filtrate centrifuging and taking lower sediment is seeded to adhere-wall culture in DMEM high glucose medium after filtering；Using nature method of purification and repeatedly adherent method purifying cells up to rheumatoid arthritis synovial fibroblast.It can get 10 using of the invention cultural method 30 days10A or so RASFs, cell yield greatly improve high, can satisfy kinds of experiments needs, therefore be a kind of quickly and effectively primary RASFS isolated culture method.Originally culture obtains RASFs and compares with the synovial cell system for being is built in vitro simultaneously, has higher researching value, the prospect with good practical application.
Description
Technical field
The invention belongs to technical field of cell culture, and in particular to a kind of rheumatoid arthritis synovial is fibroblastic
Primary culture method.
Background technique
Disclosing the information of the background technology part, it is only intended to increase understanding of the overall background of the invention, without certainty
It is considered as recognizing or implying in any form that information composition has become existing skill well known to persons skilled in the art
Art.
Rheumatoid arthritis (Rhe μm of atoid arthritis, RA) is a kind of to be related to the autoimmune disease of whole body.
The major pathologic features of RA are arthritis synovial chronic inflammatory reactions, inflammatory cell infiltration, and pannus is formed, and bone and cartilage carry out
Property destroy.The hyperplasia of synovial membrane and the violent inflammatory reaction of recurrent exerbation lead to the destruction of joint joint bone and cartilage, cause
Joint wear, joint function disturbance.Although people have been achieved for certain progress to the treatment of RA in recent years, it is specific
The cause of disease is still unclear.RASFs is directed to patient's lesions position, plays in RA patient bone and cartilage destruction important
Effect.A large amount of inflammatory cell infiltration, which is proliferated RASFs largely, in patient part causes bone and cartilage progressive to destroy.Closely
It is higher using valence to confirm that the related genes expression in regulation RASFs has in RA diagnosing and treating for numerous studies over year
Value.
Therefore, at present in the Effect study in RA pathogenesis, RASFs becomes the important cells of synovial membrane functional study
One of model.The primary culture method of RASFs mainly includes tissue mass cell culture and mixing enzyme digestion at present.Uterus tissue pieces
Method is easy to fall off since the attaching of tissue block and cell culture vessel is not firm, and cultivation cycle is long, and digestion time is not easily-controllable
System, easily pollutes, success rate is not high.And mix enzyme digestion and mix pancreatin and clostridiopetidase A, tissue digestion is carried out, due to pancreatin
It is larger to cell membrane damage, and synovial cell is detached from the constraint of tissue, climbs out of from tissue block, needs longer digestion time,
Very big to the damage of cell membrane, the cell state of acquisition is often bad.Although previous research carries out synovial membrane by mechanical shearing
Certain is broken, is then digested using mixing enzyme digestion, improves the acquisition efficiency of primary RASFs to a certain extent,
But inventor has found two main problems during the experiment: 1, RA tissue is based on fibroid tissue, and clostridiopetidase A can
Very effective digestion fibroid tissue, and damage of the pancreatin to cell is very big, and prolonged digestion will lead to a large amount of thin
Born of the same parents are dead.2, original extraction scheme does not quantify the volume of synovial tissue's block, easily makes the excess or deficiency of enzyme,
Cause cell death or digestion insufficient, final primary RASFs extraction efficiency is low.
Summary of the invention
Based on above-mentioned the deficiencies in the prior art, the present invention provides a kind of fibroblastic original of rheumatoid arthritis synovial
For cultural method, the present invention is by optimizing conventional primary synovial cell's extracting method, to greatly improve synovial cell
Extraction efficiency can obtain many experiments sample in a short time, and only not subsequent rheumatoid arthritis study of incident mechanism is established
Basis is determined, and experimental period can be greatly shortened.And originally culture obtains RASFs and builds the synovial cell system phase for being in vitro
Than having higher researching value, therefore the value with good practical application.
One aspect of the present invention provides a kind of fibroblastic primary culture method of rheumatoid arthritis synovial,
The described method includes:
Synovial tissue after pretreatment is sufficiently broken；III Collagenase Type and II Collagenase Type are added thereto and carries out
Piping and druming；
Individual cells are dispersed as after culture digestion；
Filtrate centrifuging and taking lower sediment is seeded to adhere-wall culture in DMEM high glucose medium after filtering；
Using nature method of purification and repeatedly adherent method purifying cells up to rheumatoid arthritis synovial fibroblast.
The second aspect of the invention provides the rheumatoid arthritis synovial of above-mentioned cultural method acquisition into fiber finer
Born of the same parents.
Advantageous effects of the invention:
The present invention provides a kind of fibroblastic primary culture method of rheumatoid arthritis synovial, using of the invention
It can get 10 within cultural method 30 days10A or so RASFs is 7-10 times of conventional digestion method, and cell yield greatly improves, and thin
Fuller in born of the same parents' form, activity is stronger, can satisfy kinds of experiments needs, therefore be a kind of quickly and effectively primary RASFS
Isolated culture method.Originally culture obtains RASFs and compares with the synovial cell system for being is built in vitro simultaneously, has higher research valence
Value, the prospect with good practical application.
Detailed description of the invention
The Figure of description for constituting a part of the invention is used to provide further understanding of the present invention, and of the invention shows
Examples and descriptions thereof are used to explain the present invention for meaning property, does not constitute improper limitations of the present invention.
Fig. 1 is that inverted microscope observes original method and optimization method of the present invention in cellular morphology in the embodiment of the present invention 1
With quantitative disparity map.
Fig. 2 is original extracting method and optimization method cell extraction quantity variance figure of the present invention in the embodiment of the present invention 1.
Fig. 3 is third generation RAFLs cell purity figure prepared by the flow cytometry identification embodiment of the present invention 1.
Specific embodiment
It is noted that described further below be all exemplary, it is intended to provide further instruction to the present invention.Unless another
It indicates, all technical and scientific terms that the present invention uses have logical with general technical staff of the technical field of the invention
The identical meanings understood.
It should be noted that term used herein above is merely to describe specific embodiment, and be not intended to restricted root
According to the illustrative embodiments of the present patent application.As used herein, unless the context clearly indicates otherwise, otherwise singular
Form be also intended to include plural form, additionally, it should be understood that, when in the present specification use term "comprising" and/or
When " comprising ", existing characteristics, step, operation, device, component and/or their combination are indicated.
As previously mentioned, conventional primary synovial cell's extracting method there are extraction efficiencies lower, primary synovial cell of acquisition
The defects of negligible amounts.
In view of this, in an exemplary embodiment of the invention, a kind of rheumatoid arthritis synovial is provided into fiber
The primary culture method of cell, which comprises
S1, synovial tissue after pretreatment is sufficiently crushed；III Collagenase Type and II Collagenase Type are added thereto；
Individual cells are dispersed as after S2, culture digestion；
Filtrate centrifuging and taking lower sediment is seeded to adhere-wall culture in DMEM high glucose medium after S3, filtering；
S4, using nature method of purification and repeatedly adherent method purifying cells up to rheumatoid arthritis synovial at fiber finer
Born of the same parents.
In another exemplary embodiment of the invention, the preprocess method includes the synovial membrane that will be soaked in physiological saline
Tissue is taken out and is rinsed under aseptic conditions using PBS, and fat and cartilage in synovial tissue are removed；
In another exemplary embodiment of the invention, breaking method is to shred, and is further preferably shredded to < 0.5mm；
In another exemplary embodiment of the invention, the concentration of III Collagenase Type be preferably 0.2~0.6 μ g/ml (preferably
0.5 μ g/ml), the concentration of II Collagenase Type is preferably 10~30 μ g/ml (preferably 20 μ g/ml)；Pass through optimum choice clostridiopetidase A
Matched proportion density, be conducive to digestion of the clostridiopetidase A to fibroid tissue, while avoiding using pancreatin, thus reduce to synovial cell
Damage；
In another exemplary embodiment of the invention, the area volume of synovial tissue and III Collagenase Type and II Collagenase Type
Than for 0.3~0.6cm2: the μ l of 2~3ml:200~400 (preferably 0.5cm2: 3ml:300 μ l)；By limiting synovial tissue and glue
Dosage relation between protoenzyme, thus avoid enzyme add it is excessive cause cell death or enzyme dosage it is very few cause to digest it is insufficient
Phenomenon occurs；
In another exemplary embodiment of the invention, blown and beaten repeatedly after III Collagenase Type and II Collagenase Type are added thereto
2~3min；It is mixed by piping and druming, comes into full contact with enzyme and synovial tissue to improve digestive efficiency；
In another exemplary embodiment of the invention, the culture digestion method particularly includes: be placed in 37 DEG C, 5%CO2Cell
3~6h of incubator digestion culture；
In another exemplary embodiment of the invention, being dispersed as individual cells specific method is using piping and druming mode, the party
Method is more soft slow, reduces the damage to cell；
In another exemplary embodiment of the invention, also there is 1% dual anti-and 10%FBS in DMEM high glucose medium；
In another exemplary embodiment of the invention, the rheumatoid arthritis synovial fibroblast is commissioned to train for 3-6
Cell is supported, it is higher to cultivate cell activity at this time.
In another exemplary embodiment of the invention, provide rheumatoid arthritis synovial that above-mentioned cultural method obtains at
Fibrocyte.
Explanation is further explained to the present invention by the following examples, but is not construed as limiting the invention.It should be understood that
These examples are only for illustrating the present invention and are not intended to limit the scope of the present invention.
Embodiment 1
1 main agents instrument consumptive material
1.1 reagent
| Reagent name | Producer | The place of production |
| Australia fetal calf serum | Gibco | The U.S. |
| II Collagenase Type | Suo Laibao | Beijing |
| III Collagenase Type | Suo Laibao | Beijing |
| DMEM cell culture medium | Hyclone | The U.S. |
| Pancreatin, penicillin streptomycin | Suo Laibao | Beijing |
| FITC marks vimentin antibody | eBiosecience | The U.S. |
| CD68 antibody | eBiosecience | The U.S. |
1.2 instrument consumptive materials
2 methods
The fibroblastic separation of 2.1 rheumatoid arthritis synovials and culture
1) cunning that the audit of our unit's ethics obtains the RA patient cut off through replacement knee in arthroplasty is agreed to and passed through through patient
Membrane tissue；
2) synovial tissue removed is placed in sterile physiological saline, is sealed in sterile sample generation and is transferred quickly to
Laboratory；
3) RA patient's knee synovial tissue of joint 20ml PBS is rinsed 3 times in superclean bench, it is every all over 2 clocks；
4) the careful fat and cartilaginous tissue removed in synovial tissue is cut using Sterile ophthalmic；
5) by synovial tissue with every small culture dish (30mm) 0.5cm2Synovial tissue averagely assigns in several capsules；
6) < 0.5mm is sufficiently shredded to the synovial tissue in every capsule using eye scissors；
7) in the tissue that every ware shreds be added 3ml (0.5 μ g/ml) III Collagenase Type and 300 μ l (20 μ g/ml) II
Collagenase Type；
8) blowing and beating (2 minutes) repeatedly using sterile pipette tips comes into full contact with enzyme and synovial tissue to improve digestive efficiency；
9) 37 DEG C are placed in, 5%CO2In cell incubator, 3-6h is digested；
10) cell is blown and beaten repeatedly using 1ml liquid-transfering gun after digesting and becomes individual cells, if can not will organize
It dispels and is put into incubator and continues to digest；
11) synovial tissue for finishing and dispelling using 70 μm of cell screen filtering digestion；
12) filtrate is moved in sterile 5ml centrifuge tube, 5min is centrifuged with 1500r/min revolving speed in centrifuge；
13) lower sediment is collected after being centrifuged to be inoculated in containing 1% dual anti-and 10%FBS DMEM high glucose medium
In；
14) RASFs can be completely adherent after 48h, carries out changing the non-attached cell of liquid removing to cell；
15) it is carried out changing liquid and passage according to cell state, using nature method of purification and adherent method purifying cells repeatedly, takes 3-
The good cell of 6 generation activity high-purities is tested.
The identification of 2.2 synovioblasts
2.1.1 morphologic observation
The primary cell form of the culture first generation and the second generation is observed using inverted microscope.Usual RASFs be synovial membrane at
Fibroblast-like cell.Whether the form for observing RASFs cell is spindle shape, whether adherent growth carrys out the preliminary purity for judging RASFs
And growth conditions.
2.1.2 cell count
Primary cell is counted using cell counter.It weighs the tissue of phase homogenous quantities by conventional method and optimizes
After method is extracted, it is inoculated into 25cm2Tissue Culture Flask in, then by nature propagating method purifying cells, finally use pancreas
Enzymic digestion cell counts cell using cell counter.
2.1.3 flow cytometry
Third generation RASFs is used into collection 2 × 10 after pancreatin digestion6Cell is added after cell is resuspended in 200 μ LPBS and is divided into
It two parts, is separately added into 20 μ L FITC and anti-human vimentin antibody and anti-human CD68 antibody is marked to mix, 4 DEG C are protected from light incubation
30min, during which it is primary to mix cell for every 5min oscillation.After washing cell twice with PBS, 500 μ L fixers is added, cell is resuspended
It is protected from light, carries out Flow cytometry as early as possible.
3 results
3.1 cellular morphology
Primary cell through 36h it is adherent after, change liquid to cell to remove heteroproteose cell, observed using inverted microscope.
General extraction methods and optimization method can efficiently extract into the RASFs of threadiness, (second generation) RASFs body after passage
Product increases, and more stretches, has had fibroblastic characteristic feature, adherence quality is more preferable.But with general extraction methods phase
More apparent than the RASFs profile that the method for the present invention is extracted, growth activity is stronger (Fig. 1).
3.2 cell count
After being digested to third generation cell, the PBS of same volume is added, is counted using cell counting board, counts
Process carries out in three times, records and counts cell quantity (Fig. 2).
3.3 flow cytometries identify RASFs purity
Originally culture third generation RASFs cell is respectively with after resisting anti-vimentin and 8 antibody of anti-CD 6 to mark, flow cytometer showed
Vimentin is positive in display gained synovial cell, cells ratio of CD68 feminine gender is up to 95% or more (Fig. 3).
The present invention is shredded synovial tissue using mechanical system as far as possible, and shreds synovial membrane to what is be added in each digestion system
Tissue is quantified, and has been abandoned former pancreatin and clostridiopetidase A mixture slaking method, has directly been used the collagen of certain volume and concentration
Enzyme mixation is digested, and is blown and beaten and mixed so as to organize to come into full contact with enzyme, raising digestive efficiency with rifle.Make after digestion
Tissue block is gently dispelled with pipette tips, uses strainer filtering.The cellular damage that this soft slow method obtains is minimum.With original method
Fuller in cellular morphology compared to the primary RASFs that the method after optimization is extracted, activity is stronger, and cell quantity is more
More, extraction efficiency is higher.Cell count also illustrates that the method after optimization extracts cell quantity more multiple-effect than original method again
Rate is higher.Vimenin is fibroblastic protein marker, using identification fibroblast marker molecule vimentin and
The method of CD68 identifies the type of obtained synovial cell.Flow cytometry shows, vimentin is positive and CD68 is negative
Cell accounts for 95% or more of all synovial cells, and original method and optimization method vimentin positive rate difference are little.With
Above the result shows that, the method for optimization can efficiently extract RASFs, and extraction efficiency greatly improves.
Finally it should be noted that the foregoing is only a preferred embodiment of the present invention, it is not limited to this hair
It is bright, although the present invention is described in detail referring to the foregoing embodiments, for those skilled in the art, still
It can modify to technical solution documented by previous embodiment, or part is equivalently replaced.It is all in this hair
Within bright spirit and principle, any modification, equivalent replacement, improvement and so on should be included in protection scope of the present invention
Within.Above-mentioned, although specific embodiments of the present invention have been described, and it is not intended to limit the protection scope of the present invention, institute
Category field technical staff should be understood that based on the technical solutions of the present invention those skilled in the art do not need to pay wound
The various modifications or changes that the property made labour can be made are still within protection scope of the present invention.
Claims (10)
1. a kind of fibroblastic primary culture method of rheumatoid arthritis synovial, which comprises
Synovial tissue after pretreatment is sufficiently broken；III Collagenase Type and II Collagenase Type are added thereto；
Individual cells are dispersed as after culture digestion；
Filtrate centrifuging and taking lower sediment is seeded to adhere-wall culture in DMEM high glucose medium after filtering；
Using nature method of purification and repeatedly adherent method purifying cells up to rheumatoid arthritis synovial fibroblast.
2. a kind of primary culture method as described in claim 1, which is characterized in that the preprocess method includes that will be soaked in
Synovial tissue in physiological saline takes out is rinsed using PBS under aseptic conditions, removes the fat and cartilage in synovial tissue.
3. a kind of primary culture method as described in claim 1, which is characterized in that breaking method is to shred, further preferably
It shreds to < 0.5mm.
4. a kind of primary culture method as described in claim 1, which is characterized in that the concentration of III Collagenase Type is preferably 0.2
~0.6 μ g/ml (preferably 0.5 μ g/ml), the concentration of II Collagenase Type are preferably 10~30 μ g/ml (preferably 20 μ g/ml).
5. a kind of primary culture method as described in claim 1, which is characterized in that synovial tissue and III Collagenase Type and II type
The area-volume ratio of clostridiopetidase A is 0.3~0.6cm2: the μ l of 2~3ml:200~400 (preferably 0.5cm2: 3ml:300 μ l).
6. a kind of primary culture method as described in claim 1, which is characterized in that III Collagenase Type and II type are added thereto
2~3min is blown and beaten after clostridiopetidase A repeatedly.
7. a kind of primary culture method as described in claim 1, which is characterized in that the culture digestion method particularly includes: set
In 37 DEG C, 5%CO23~6h of cell incubator digestion culture；
The individual cells specific method that is dispersed as is to be carried out using piping and druming mode.
8. a kind of primary culture method as described in claim 1, which is characterized in that also have in DMEM high glucose medium
1% dual anti-and 10%FBS.
9. a kind of primary culture method as described in claim 1, which is characterized in that the rheumatoid arthritis synovial is at fibre
Dimension cell is 3-6 subtituted culturing cell.
10. the rheumatoid arthritis synovial fibroblast that any one of the claim 1-9 primary culture method obtains.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910458872.5A CN110117570B (en) | 2019-05-29 | 2019-05-29 | Primary culture method of rheumatoid arthritis synovial fibroblasts |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910458872.5A CN110117570B (en) | 2019-05-29 | 2019-05-29 | Primary culture method of rheumatoid arthritis synovial fibroblasts |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110117570A true CN110117570A (en) | 2019-08-13 |
| CN110117570B CN110117570B (en) | 2020-03-10 |
Family
ID=67523514
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910458872.5A Expired - Fee Related CN110117570B (en) | 2019-05-29 | 2019-05-29 | Primary culture method of rheumatoid arthritis synovial fibroblasts |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110117570B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111411076A (en) * | 2020-03-24 | 2020-07-14 | 山东省医药生物技术研究中心（山东省病毒研究所） | Method for efficiently separating primary cells of human synovial membrane |
| CN111471642A (en) * | 2020-03-24 | 2020-07-31 | 山东省医药生物技术研究中心（山东省病毒研究所） | Method for efficiently promoting synovial cell proliferation based on tension effect |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101708328A (en) * | 2009-11-06 | 2010-05-19 | 上海市免疫学研究所 | Pharmaceutical application of CYR61 protein |
| CN101747435A (en) * | 2008-12-18 | 2010-06-23 | 上海市免疫学研究所 | Monoclonal antibody (hybrid tumor) for neutralizing CYR61 and applications thereof |
-
2019
- 2019-05-29 CN CN201910458872.5A patent/CN110117570B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101747435A (en) * | 2008-12-18 | 2010-06-23 | 上海市免疫学研究所 | Monoclonal antibody (hybrid tumor) for neutralizing CYR61 and applications thereof |
| CN101708328A (en) * | 2009-11-06 | 2010-05-19 | 上海市免疫学研究所 | Pharmaceutical application of CYR61 protein |
Non-Patent Citations (4)
| Title |
|---|
| JACQUES MOREL等: "Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) induces rheumatoid arthritis synovial fibroblast proliferation through mitogen-activated protein kinases and phosphatidylinositol 3-kinase/Akt", 《THE JOURNAL OF BIOLOGICAL CHEMISTRY》 * |
| 丁娟等: "RA滑膜成纤维细胞的原代培养及鉴定", 《现代生物医学进展》 * |
| 仇海文等: "II型胶原诱导的关节炎大鼠滑膜成纤维细胞的培养、纯化和鉴定", 《南京师大学报（自然科学版）》 * |
| 马莎等: "三种培养原代滑膜成纤维细胞方法的比较", 《昆明医科大学学报》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111411076A (en) * | 2020-03-24 | 2020-07-14 | 山东省医药生物技术研究中心（山东省病毒研究所） | Method for efficiently separating primary cells of human synovial membrane |
| CN111471642A (en) * | 2020-03-24 | 2020-07-31 | 山东省医药生物技术研究中心（山东省病毒研究所） | Method for efficiently promoting synovial cell proliferation based on tension effect |
| CN111471642B (en) * | 2020-03-24 | 2022-07-26 | 山东省医药生物技术研究中心（山东省病毒研究所） | Method for efficiently promoting synovial cell proliferation based on tension effect |
| CN111411076B (en) * | 2020-03-24 | 2022-09-23 | 山东省医药生物技术研究中心（山东省病毒研究所） | Method for efficiently separating primary cells of human synovial membrane |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110117570B (en) | 2020-03-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106754674B (en) | The method and its application of amnion mesenchymal stem cell are prepared from Human plactnta amnion | |
| Jonason et al. | Primary murine growth plate and articular chondrocyte isolation and cell culture | |
| JP6687757B2 (en) | Methods for preparing 3D cartilage organoid blocks | |
| Häkkinen et al. | An improved method for culture of epidermal keratinocytes from newborn mouse skin | |
| CN101765657A (en) | Method for isolating and culturing adult stem cells derived from human amniotic epithelium | |
| CN106801032B (en) | Construction method of human amniotic epithelial stem cell bank | |
| CN105238749B (en) | A kind of method of recovery mesenchymal stem cell | |
| CN104498433A (en) | Extraction method of adipose-derived stem cells as well as preparation and application of adipose-derived stem cells | |
| CN105200007B (en) | The method that Subaerial blue green algae is extracted from placental fetal surface chorion | |
| CN105087474A (en) | Culture method of deciduous tooth pulp stem cells | |
| CN107921182A (en) | Mechanical device and method for isolation medium vasculature part | |
| CN110117570A (en) | A kind of fibroblastic primary culture method of rheumatoid arthritis synovial | |
| CN106011055A (en) | Preparation method of human primary cartilage cells with high yield rate | |
| CN105441391A (en) | Cell model for diabetes complicated with depression, and establishing method and application thereof | |
| CN113322231A (en) | Method for separating and culturing mesenchymal stem cells and preparation | |
| CN104031881B (en) | A kind of isolated culture method of mankind's olfactory mucosa mescenchymal stem cell | |
| CN107227296A (en) | A kind of method of myelomonocyte separation, identification and induction differentiation | |
| CN108486039A (en) | The method that small molecule induction human adipose-derived stem cell is divided into interstitial glands | |
| CN106834232A (en) | Pituitary adenoma cell system and application thereof | |
| CN109988743A (en) | A method of rapidly and efficiently separating Human plactnta amniotic epithelial cells | |
| CN104818243A (en) | Separation method of placenta-derived fetal stem cells | |
| CN113025661A (en) | Construction method of immortalized musk glandular epithelial cells | |
| CN102424817B (en) | Cell strain from human lung adenocarcinoma and preparation method thereof | |
| CN108774630A (en) | A kind of primary culture method of Microhyla ornata Skeletal Muscle Cell | |
| CN115029305A (en) | Separation and identification method for pig FAPs (FAPs) cells and application of FAPs 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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20200310 Termination date: 20200529 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |