CN108676771A - The separation method and separating obtained PMCs of a kind of primary Peritoneal Mesothelial Cells - Google Patents
The separation method and separating obtained PMCs of a kind of primary Peritoneal Mesothelial Cells Download PDFInfo
- Publication number
- CN108676771A CN108676771A CN201810521528.1A CN201810521528A CN108676771A CN 108676771 A CN108676771 A CN 108676771A CN 201810521528 A CN201810521528 A CN 201810521528A CN 108676771 A CN108676771 A CN 108676771A
- Authority
- CN
- China
- Prior art keywords
- pmcs
- primary
- centrifugation
- cell
- patient
- 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
-
- 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
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Zoology (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)
Abstract
The present invention relates to a kind of separation method of primary Peritoneal Mesothelial Cells and separating obtained PMCs.The method of the invention detaches primary PMCs from the dialyzate of the patient in after peritoneal dialysis tube placing operation 2 weeks.The method step is simple, noninvasive and be easy to repeat, and the primary PMCs of gained has many advantages, such as that purity is high, adherence quality is good, passage number is more, amalgamation is good, can directly reflect physiological changes of the PD patient PMCs in PDS.
Description
Technical field
The present invention relates to field of cell culture, a kind of separation method in particular to primary Peritoneal Mesothelial Cells and
Separating obtained PMCs.
Background technology
Peritoneal Mesothelial Cells (peritoneal mesothelial cells, PMCs) are to constitute the most important cell of peritonaeum
Group is both a kind of biological barrier and a secretory, can synthesize and secrete various substances, is maintaining peritonaeum stable state, object
It plays a significant role in matter transhipment, inflammation and immunological regulation, tissue repair, and PMCs is directly long with dialyzate in dialysis procedure
Time contacts, biological barriers of the PMCs as peritonaeum, carries out the transhipment of ultrafiltration and solute, is maintaining in peritonaeum in the stabilization of environment
It plays a crucial role.Therefore, the primary PMCs of in vitro culture inquires into the pathophysiological process of peritonaeum, fine for prevention peritonaeum
The generation of dimensionization, extension peritonaeum service life are of great significance.
The cultural method of current common primary PMCs is detached from the peritoneal tissues of surgery patients, such as greatly
Nethike embrane, but this method has that materials are difficult and are related to Medicine Ethics.Another method is the dialysis stream from PD patient
Go out and obtains PMCs in liquid, it is adherent slow compared with the primitive cell culture from omentum majus although this method is simple, and pass
In generation, is few, and cell will lose vigor after being generally passaged to for the 4th generation, cannot merge.
In view of this, special propose the present invention.
Invention content
The first object of the present invention is to provide a kind of separation method of primary Peritoneal Mesothelial Cells, the method step letter
Single, noninvasive and be easy to repeat, the primary PMCs of gained has that purity is high, adherence quality is good, passage number is more, amalgamation is good, can be direct
The advantages that reflecting physiological changes of the PD patient PMCs in PDS.
The second object of the present invention is to provide a kind of primary PMCs obtained according to preceding method, the primary PMCs tools
Have that purity is high, adherence quality is good, passage number is more, amalgamation is good, can directly reflect physiological changes etc. of the PD patient PMCs in PDS
Advantage.
In order to realize that the above-mentioned purpose of the present invention, spy use following technical scheme：
A kind of separation method of primary Peritoneal Mesothelial Cells (peritoneal mesothelial cells, PMCs), institute
It states method and detaches PMCs from the dialyzate of the patient in after peritoneal dialysis tube placing operation 2 weeks.
The invention further relates to the primary PMCs obtained according to preceding method.
Specific implementation mode
The present invention relates to a kind of points of primary Peritoneal Mesothelial Cells (peritoneal mesothelial cells, PMCs)
From method, the method detaches primary PMCs from the dialyzate of the patient in after peritoneal dialysis tube placing operation 2 weeks.
Primary PMCs usually from peritoneal tissues (such as omentum majus) or from the dialysis efflux of general PD patient, the former
Not only materials difficulty further relates to medical ethics problems, and the separating obtained cell of the latter is adherent slow, and passage is few, is generally passaged to the 4th
It will lose vigor, cannot merge for later cell.
In order to solve the above technical problems, the present invention provides the separation method of aforementioned primary peritoneal cell, the method from
Isolated PMCs in the Peritoneal dialysate of patient after peritoneal dialysis tube placing operation in 2 weeks.The method of the invention separation
The patient of pipe operation is set in row peritoneal dialysis in the near future in the sources PMCs, and first, the method for the invention step is simple, noninvasive and be easy to
It repeats, can be that patient reduces pain；Secondly, the PMCs that the method for the invention obtains can directly reflect patients undergoing peritoneal dialysis
Physiological changes of the PMCs in peritoneal dialysis solution accomplishes fluently solid foundation for next step in vitro study peritonaeum pathologic, physiologic；Furthermore
The PMCs of the method for the invention be derived from recent row peritoneal dialysis set pipe operation patient, positioned at the PMCs in the stage can retain compared with
Good functional status, is conducive to the research of its pathophysiological, and process is the study found that primary PMCs trainings described in the method for the invention
It supports to the 10th generation, gives 1：When 3 ratios are passed on, cell can merge after 1-2 days, show good fusion faculty.
In some specific embodiments, the method specifically includes：The dialyzate of patient described in sterile collection, through from
After the heart and washing step, cell is resuspended up to the primary PMCs using culture medium.
In some specific embodiments, the centrifugation step includes：The dialyzate is dispensed into centrifuge tube,
At 3~10 DEG C, 300~500g centrifuges 5~15min.
Preferably, described centrifuge includes：The dialyzate is dispensed into centrifuge tube, at 4 DEG C, 400g centrifuges 10min.
In some specific embodiments, the washing step includes：After the centrifugation step, supernatant is abandoned, with slow
Centrifugation gained sedimentation cell is resuspended in fliud flushing, and is centrifuged again.
In some specific embodiments, the washing step further includes：Supernatant is abandoned again after centrifugation described,
Centrifugation gained sedimentation cell is resuspended with buffer solution, and carries out third time centrifugation.
In some specific embodiments, the condition of the centrifugation again and/or third time centrifugation is：3~
At 10 DEG C, 300~500g centrifuges 5~15min.
In some specific embodiments, the condition of the centrifugation again and/or third time centrifugation is：At 4 DEG C
Under, 400g centrifuges 10min.
In some specific embodiments, the culture medium is complete medium, it is preferable that the complete medium is
The DMEM/F12 culture mediums of FBS containing 10%v/v, 1%w/v penicillin and 1%w/v streptomysins.
In some specific embodiments, the method further includes：Cultivate the primary PMCs cells；Preferably, institute
Stating culture includes：The primary PMCs is placed in 37 DEG C, 5%CO2Under conditions of cultivated, liquid was changed every 2~3 days 1 time.
In some specific embodiments, the method further includes being passed on to the primary PMCs, it is preferable that
The growth of cell is passed on when being fused to 80~90%.
In some specific embodiments, the method further includes that the primary PMCs is frozen and/or recovered.
In some specific embodiments, the patient is the End-stage renal disease in row peritoneal dialysis tube placing operation 2 weeks
(End stage renal disease, ESRD) patient.
The invention further relates to the primary PMCs obtained according to preceding method.
Compared with prior art, beneficial effects of the present invention are：
The primary PMCs that the present invention provides a kind of separation method of primary Peritoneal Mesothelial Cells and obtained by this method, institute
It is simple, noninvasive and be easy to repeat to state method and step, the primary PMCs of gained has that purity is high, adherence quality is good, passage number is more, merges
Property is good, the advantages that capable of directly reflecting physiological changes of the PD patient PMCs in PDS.
Description of the drawings
It, below will be to specific in order to illustrate more clearly of the specific embodiment of the invention or technical solution in the prior art
Embodiment or attached drawing needed to be used in the description of the prior art are briefly described, it should be apparent that, in being described below
Attached drawing is some embodiments of the present invention, for those of ordinary skill in the art, before not making the creative labor
It puts, other drawings may also be obtained based on these drawings.
The Morphological Features (experimental example 1) that Fig. 1 is primary PMCs described in embodiment 1 under inverted microscope, wherein：Figure 1A is P0
Generation, Figure 1B P4The amplification factor in generation, Figure 1A~Figure 1B is 100 times；
Fig. 2 is the immunofluorescence dyeing result (experimental example 2) of primary PMCs described in embodiment 1, wherein Fig. 2A is CK18's
Coloration result, Fig. 2 B are the coloration result of vimentin, and Fig. 2 C are the coloration result of DAPI, and Fig. 2 D are Fig. 2A~Fig. 2 C's
Merge image results.
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will
Understand, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the invention.It is not specified in embodiment specific
Condition person carries out according to conventional conditions or manufacturer's recommended conditions.Reagents or instruments used without specified manufacturer is
The conventional products obtained can be bought by city.
Embodiment 1
The present embodiment provides the separation of primary PMCs a kind of and cultural method, the method to specifically include following steps：
1, the separation of primary PMCs
(1) ESRD patient at night stays the peritoneal effluent sample (PDS) of abdomen in 2 weeks after sterile collection peritoneal dialysis tube placing operation；
(2) PDS is dispensed into 250ml centrifuge tubes on steril cell operation console, 4 DEG C of 400g are centrifuged 10 minutes；
(3) supernatant is abandoned, after sedimentation cell adds PBS to be resuspended, absorption moves to 15ml sterile centrifugation tubes, in 4 DEG C, the condition of 400g
Under, it centrifuges 5 minutes；
(4) step (3) is repeated；
(5) supernatant is abandoned, is resuspended with appropriate complete medium (DMEM/F12 containing 10%FBS and 1% penicillin/streptomycin)
Cell, the complete medium are FBS containing 10%v/v, 1%wt penicillin and 1%wt streptomysin DMEM/F12 culture mediums.
2, the culture of primary PMCs
(1) cell suspension is moved in 75ml culture bottles, adjustment complete medium is 20ml；
(2) culture bottle is put to 37 DEG C, 5%CO2Cell incubator in cultivate；
(3) it was changed the liquid once every 2-3 days.
Embodiment 2
The present embodiment provides the propagating method of aforementioned primary PMCs, the method specifically includes following steps：
(1) it is passed on when cell growth is fused to 80-90%；
(2) 75cm is absorbed2Old culture medium in gas-permeable flasks is added 2ml PBS, gently shakes, wash away remaining training
Support base and suspension cell；
(3) 0.25% pancreatin of 1.5ml (containing 0.02%EDTA) covering bottom of bottle is added, is then placed in 37 DEG C of incubators and incubates
About 1min is educated, cell dissociation situation is dynamically observed under microscope；
(4) show that the DMEM/F12 containing 10%FBS is added when cell rounding does not float terminates digestion in right amount under microscope；
(5) it gently blows and beats into be transferred in 15ml centrifuge tubes after cell suspension and centrifuge, 4 DEG C, 1000rpm, centrifuge 5min；
(6) supernatant is abandoned, is gently blown and beaten with culture medium, cell suspension is formed, with 1:In 3 packing to new culture bottles, it is placed in thin
Continue to cultivate in born of the same parents' incubator.
Embodiment 3
The present embodiment provides aforementioned primary PMCs freeze and method for resuscitation, the method specifically include following steps：
1, cell cryopreservation
(1) when cell growth is fused to about 80-90%, culture medium old in culture bottle is sucked, 2mlPBS is added, gently
It shakes, washes away remaining culture medium and suspension cell；
(2) 1.5ml pancreatin is added, puts into 37 DEG C of incubators and is incubated about 1min, cell dissociation is dynamically observed under microscope
Situation；
(3) 2ml containing complete medium is added when cell rounding does not float and terminates digestion；
(4) it gently blows and beats into be transferred in 15ml centrifuge tubes after cell suspension and centrifuge, 4 DEG C, 1000rpm, centrifuge 5min；
(5) supernatant is abandoned, frozen stock solution (DMEM/F12 is added：FBS:DMSO=7：2：1) cell is resuspended, packing is moved into and frozen
Pipe, is put into -80 DEG C of refrigerators after being placed in gradient cooling box, is transferred in liquid nitrogen and preserves in 1 week.
2, cell recovery
(1) PMCs cells are taken out from liquid nitrogen container, are immediately placed in 37 DEG C of water baths, are shaken frequently；
(2) 75% cotton ball soaked in alcohol wipe cryopreservation tube nozzle, uncap in super-clean bench, and it is complete to 5ml is equipped with that cell suspension is sucked out
In the 15ml centrifuge tubes of culture medium, 4 DEG C, 1000rpm, 5min is centrifuged；
(3) supernatant is abandoned, is gently blown and beaten with complete medium, cell suspension is formed and moves to 75cm2In gas-permeable flasks, set
In 37 DEG C, 5%CO2It is cultivated in cell incubator；
(4) it gently blows and beats into be transferred in 15ml centrifuge tubes after cell suspension and centrifuge；
(5) culture medium is replaced afterwards for 24 hours to continue to cultivate.
Experimental example 1
The cellular morphology of primary PMCs described in Examples 1 to 2 is observed under inverted microscope, wherein testing result is as follows
It is shown：
The PMCs of visible original cuiture initially performances are mostly starlike or fusiform, the strong (figure of translucency under inverted microscope
1A)；Cell growth is fused to about 80-90% after 1 week, can pass down；3-4 for when, cell fusion at when be in polygon, shape
At typical paving stone shape (Figure 1B).
Experimental example 2
To further determine that whether 1 Isolated cells of embodiment are PMCs, 1 Isolated cells of embodiment are exempted from
Epidemic disease fluorescent staining and microscopy.Testing result according to Fig.2, it is found that 1 Isolated cells of the embodiment of the present invention cell born of the same parents
It is the positive to starch CK18 and vimentin, it was demonstrated that 1 Isolated cells of the embodiment of the present invention are PMCs.
Immunofluorescent staining and observation are shown in the specific method is as follows：
(1) routine passage cell is pre-placed the coverslip for being coated with gelatin to 24 orifice plates in culture dish；
(2) when cell fusion is at single layer, culture medium is sucked, 1 × PBS is rinsed 3 times；
(3) 4% poly methanol is added, room temperature fixes 10min；
(4) 1 × PBS are washed 3 times；
(5) 0.1%Triton is added, room temperature is incubated 10min；
(6) 1 × PBS are washed 3 times；
(7) while the monoclonal antibody (1 of the anti-CK18 of mouse is added:25in 1%BSA) and rabbit-anti Vimentin monoclonals it is anti-
Body (1:25in 1%BSA), 4 DEG C of overnight incubations；
(8) primary antibody is sucked, 1 × PBS is washed 3 times；
(9) while Donkey anti-Mouse and the Donkey anti-that Alexa 488 and Alexa 546 is marked is added
Rabbit IgG(1：1000in 1%BSA) about 100ul, room temperature incubation 1h；
(10) secondary antibody is sucked, 1 × PBS is washed 3 times, each 5min (being protected from light)；
(11) DDW is washed 1 time；
(12) DAPI redyes core：1:100in DDW, room temperature, 3min；
(13) DDW is washed 1 time；
(14) 1 drop mountant (Mounting Medium) is added dropwise in clean glass slide, coverslip is placed on glass slide
It goes up (cell is adherent down), is protected from light air dried overnight under room temperature, is observed under laser confocal microscope.
Experimental example 3
2 the method according to embodiments of the present invention presses 1 to primary PMCs described in the embodiment of the present invention 1:3 ratio is from P0Generation
Continuous passage is to P10In generation, the fusion faculty of the primary PMCs, concrete outcome are as shown in table 1 after observation passage.According to table 1
Experimental data is it is found that primary PMCs described in the embodiment of the present invention 1 has good fusion faculty：From P0It is commissioned to train and supports to P10Generation, carefully
Born of the same parents can merge after 1-2 days.
1 primary PMCs (P of table0-P10Generation) fusion faculty
| P0Generation | P1Generation | P2Generation | P3Generation | P4Generation | P5Generation | |
| Whether primary PMCs has fusion faculty | It is | It is | It is | It is | It is | It is |
| The time required to fusion (day) | 1 day | 1 day | 1 day | 1 day | 1 day | 1 day |
| P6Generation | P7Generation | P8Generation | P9Generation | P10Generation | ||
| Primary PMCs whether fusion faculty | It is | It is | It is | It is | It is | |
| The time required to fusion (day) | 2 days | 2 days | 2 days | 2 days | 2 days |
Finally it should be noted that：The above embodiments are only used to illustrate the technical solution of the present invention., rather than its limitations；To the greatest extent
Present invention has been described in detail with reference to the aforementioned embodiments for pipe, but it will be understood by those of ordinary skill in the art that：Its
It still can be with technical scheme described in the above embodiments is modified, either to which part or all technical features
Carry out equivalent replacement；And these modifications or replacements, various embodiments of the present invention skill that it does not separate the essence of the corresponding technical solution
The range of art scheme.
Claims (10)
1. a kind of separation method of primary Peritoneal Mesothelial Cells (peritoneal mesothelial cells, PMCs), special
Sign is that the method detaches primary PMCs from the dialyzate of the patient in after peritoneal dialysis tube placing operation 2 weeks.
2. according to the method described in claim 1, it is characterized in that, the method specifically includes：Patient described in sterile collection
Cell is resuspended up to the primary PMCs using culture medium through centrifuging with after washing step in dialyzate.
3. according to the method described in claim 2, it is characterized in that, the centrifugation step includes：By the dialyzate dispense to
In centrifuge tube, at 3~10 DEG C, 300~500g centrifuges 5~15min；
Preferably, described centrifuge includes：The dialyzate is dispensed into centrifuge tube, at 4 DEG C, 400g centrifuges 10min.
4. according to the method described in claim 2, it is characterized in that, the washing step includes：After the centrifugation step, abandon
Supernatant is resuspended centrifugation gained sedimentation cell with buffer solution, and is centrifuged again；
Preferably, the washing step further includes：Supernatant is abandoned again after centrifugation described, centrifugation gained is resuspended with buffer solution
Sedimentation cell, and carry out third time centrifugation；
It is highly preferred that the condition of the centrifugation again and/or third time centrifugation is：At 3~10 DEG C, 300~500g from
5~15min of the heart；
Most preferably, the condition of the centrifugation again and/or third time centrifugation is：At 4 DEG C, 400g centrifuges 10min.
5. according to the method described in claim 2, it is characterized in that, the culture medium is complete medium, it is preferable that described complete
Full culture medium is the DMEM/F12 culture mediums of FBS containing 10%v/v, 1%w/v penicillin and 1%w/v streptomysins.
6. according to the method described in claim 1, it is characterized in that, the method further includes：Cultivate the primary PMCs cells；
Preferably, described cultivate includes：The primary PMCs is placed in 37 DEG C, 5%CO2Under conditions of cultivated, every 2~3 days
Change liquid 1 time.
7. according to the method described in claim 1, it is characterized in that, the method further includes being passed to the primary PMCs
Generation, it is preferable that passed on when the growth of cell is fused to 80~90%.
8. according to the method described in claim 1, it is characterized in that, the method further includes being frozen to the primary PMCs
And/or recovery.
9. according to the method described in claim 1, it is characterized in that, the patient is the end in row peritoneal dialysis tube placing operation 2 weeks
End stage renal disease (End stage renal disease, ESRD) patient.
10. the primary PMCs obtained according to any one of claim 1~9 the method.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810521528.1A CN108676771A (en) | 2018-05-28 | 2018-05-28 | The separation method and separating obtained PMCs of a kind of primary Peritoneal Mesothelial Cells |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810521528.1A CN108676771A (en) | 2018-05-28 | 2018-05-28 | The separation method and separating obtained PMCs of a kind of primary Peritoneal Mesothelial Cells |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108676771A true CN108676771A (en) | 2018-10-19 |
Family
ID=63808533
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810521528.1A Pending CN108676771A (en) | 2018-05-28 | 2018-05-28 | The separation method and separating obtained PMCs of a kind of primary Peritoneal Mesothelial Cells |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108676771A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109810939A (en) * | 2019-04-09 | 2019-05-28 | 武汉轻工大学 | The cultural method of one boar Peritoneal Mesothelial Cells |
| CN109971685A (en) * | 2019-04-09 | 2019-07-05 | 武汉轻工大学 | A kind of construction method of haemophilus parasuis infection pig Peritoneal Mesothelial Cells model |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001002004A1 (en) * | 1999-07-02 | 2001-01-11 | Lee Hibahl | Peritoneal dialysis solution containing antioxidant for treating renal failure |
| CN107974429A (en) * | 2017-11-29 | 2018-05-01 | 宁夏医科大学总医院 | A kind of method and Optimal Medium of quick separating culture human airway epithelial cells |
| CN107988159A (en) * | 2017-12-14 | 2018-05-04 | 武汉大学深圳研究院 | A kind of method that primary tumor cell is separately cultured using malignant pleural effusion |
| CN108018258A (en) * | 2016-11-04 | 2018-05-11 | 江苏齐氏生物科技有限公司 | A kind of isolation and culture method of primary people's esophageal epithelial cell |
-
2018
- 2018-05-28 CN CN201810521528.1A patent/CN108676771A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001002004A1 (en) * | 1999-07-02 | 2001-01-11 | Lee Hibahl | Peritoneal dialysis solution containing antioxidant for treating renal failure |
| CN108018258A (en) * | 2016-11-04 | 2018-05-11 | 江苏齐氏生物科技有限公司 | A kind of isolation and culture method of primary people's esophageal epithelial cell |
| CN107974429A (en) * | 2017-11-29 | 2018-05-01 | 宁夏医科大学总医院 | A kind of method and Optimal Medium of quick separating culture human airway epithelial cells |
| CN107988159A (en) * | 2017-12-14 | 2018-05-04 | 武汉大学深圳研究院 | A kind of method that primary tumor cell is separately cultured using malignant pleural effusion |
Non-Patent Citations (2)
| Title |
|---|
| 周循等: "腹膜透析流出液中人腹膜间皮细胞的培养及转分化特征 ", 《中国现代医学杂志》 * |
| 曹东维等: "腹膜透析液对腹膜间皮细胞周期和周期蛋白的影响及骨髓间充质干细胞干预的作用 ", 《江苏医药》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109810939A (en) * | 2019-04-09 | 2019-05-28 | 武汉轻工大学 | The cultural method of one boar Peritoneal Mesothelial Cells |
| CN109971685A (en) * | 2019-04-09 | 2019-07-05 | 武汉轻工大学 | A kind of construction method of haemophilus parasuis infection pig Peritoneal Mesothelial Cells model |
| CN109971685B (en) * | 2019-04-09 | 2022-11-29 | 武汉轻工大学 | Construction method of model of porcine peritoneal epithelial cells infected by haemophilus parasuis |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| McGuire et al. | Role of extracellular matrix in regulating fenestrations of sinusoidal endothelial cells isolated from normal rat liver | |
| Jiang et al. | Reconstruction of the corneal epithelium with induced marrow mesenchymal stem cells in rats | |
| Kobayashi et al. | Corneal regeneration by transplantation of corneal epithelial cell sheets fabricated with automated cell culture system in rabbit model | |
| WO2014114043A1 (en) | Cell matrix modified tissue engineering nerve graft for repairing peripheral nerve injury and preparation method thereof | |
| CN108676771A (en) | The separation method and separating obtained PMCs of a kind of primary Peritoneal Mesothelial Cells | |
| Fang et al. | Construction of tissue-engineered heart valves by using decellularized scaffolds and endothelial progenitor cells | |
| CN111330082B (en) | Preparation method for constructing biological 3D printing skin micro-unit model containing skin accessory | |
| CN110419505A (en) | Mouse cancer of the esophagus model and its method for building up | |
| CN106916787A (en) | A kind of limbal stem cell culture medium and its cultural method | |
| CN108300688A (en) | Primary hepatocyte detaches and cultural method | |
| CN107557329A (en) | A kind of Brain Microvascular Endothelial separation and cultural method | |
| CN103230623A (en) | Method for in-vitro construction of tissue engineered nerves | |
| Nakajima et al. | A novel closed cell culture device for fabrication of corneal epithelial cell sheets | |
| Zhu et al. | Three dimensional tubular structure self-assembled by vascular mesenchymal cells at stiffness interfaces of hydrogels | |
| CN107019816A (en) | A kind of compound method for going epidermis dermis to build organization engineering skin of amniotic epithelial cells | |
| CN107034182A (en) | A kind of preparation method of fat stem cell freeze-dried powder | |
| CN108410800B (en) | It is a kind of cultivate human amnion membrane culture medium and application | |
| CN109897815A (en) | It is a kind of without coated fatty endothelial progenitor cells efficiently separate and cultural method | |
| Morris et al. | Adhesion of mouse blastocysts to uterine epithelium in culture: a requirement for mutual surface interactions | |
| CN106434530A (en) | Endothelial cell culture solution | |
| CN110317777A (en) | A kind of separation of Buccal mucosa cell and cultural method | |
| Danti et al. | Growing bone tissue-engineered niches with graded osteogenicity: an in vitro method for biomimetic construct assembly | |
| Palladini et al. | Modifications of vimentin filament architecture and vimentin–nuclear interactions by cholesterol oxides in 73/73 endothelial cells | |
| CN109913419A (en) | A kind of quadruple cynapse compound cells model, method for building up and the application of diabetes complicated depression | |
| CN107488628A (en) | A kind of simple and efficient amniotic fluid stem cell cultural method |
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: 20181019 |
|
| RJ01 | Rejection of invention patent application after publication |