CN106190952A - A kind of amplifying preparation process of epithelial cell - Google Patents
A kind of amplifying preparation process of epithelial cell Download PDFInfo
- Publication number
- CN106190952A CN106190952A CN201610569366.XA CN201610569366A CN106190952A CN 106190952 A CN106190952 A CN 106190952A CN 201610569366 A CN201610569366 A CN 201610569366A CN 106190952 A CN106190952 A CN 106190952A
- Authority
- CN
- China
- Prior art keywords
- epithelial cell
- culture fluid
- culture
- mammary gland
- preparation process
- 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
- C12N5/0625—Epidermal cells, skin cells; Cells of the oral mucosa
-
- 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
- C12N2500/30—Organic 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
- C12N2501/10—Growth factors
- C12N2501/105—Insulin-like growth factors [IGF]
-
- 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)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Dermatology (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 invention discloses the amplifying preparation process of a kind of epithelial cell, carry out in accordance with the following steps: under aseptic condition, cow mammary gland piece of tissue is washed and shredded, add trypsin solution digestion certain time, supernatant is abandoned afterwards after centrifugal for cow mammary gland tissue suspension, add resuspended rear filtration in epithelial cell culture fluid, in culture fluid inoculated and cultured ware after filtering, it is placed in incubator cultivation；Suck culture fluid after original cuiture 60 72h, be centrifuged after adding Trypsin-EDTA solution digestion and be inoculated in culture dish, being placed in incubator and carry out Secondary Culture.A kind of epithelial cell culture fluid proposed in the present invention, IGF II, prolactin antagonist, progesterone and linoleic acid is added in culture fluid, IGF II and prolactin antagonist can stimulate beta lactoglobulin and the caseic expression of β, progesterone can promote that mammary gland tissue is reached maturity, maintain gestation and lactogenic, linoleic acid can be with the biological activity of inducing mammary epithelial cell, and the culture fluid configured has facilitation to the In vitro culture of epithelial cell.
Description
Technical field
The invention belongs to technical field of cell culture, particularly relate to the amplifying preparation process of a kind of epithelial cell.
Background technology
Cell is cultivated and is referred to take out cell from in-vivo tissue, then simulated in vivo environment, at aseptic, proper temperature and suitable
Preferably acid-base value under certain nutritional condition so that it is growth and breeding, and maintain a kind of culture technique of its 26S Proteasome Structure and Function.Cell
The culture cultivated is individual cells or cell mass.The research that galactophore epithelial cell is cultivated starts to walk to compare early comparatively speaking.1961
Year, after Ebner has the most successfully cultivated cow galactophore epithelial cell the 1st time, many researcheres cultivate with cell respectively and
The mode of organ culture has cultivated galactophore epithelial cell, and carries out the influencing mechanism of milk synthesis and secretion with it for model
Research.Owing to galactophore epithelial cell is a kind of well differentiated cell, so the optimal bar of its In vitro culture must constantly be groped
Part and the effect of the interpolation factor, galactophore epithelial cell is easily separated from differentiation in incubation simultaneously, it is necessary at suitable condition
Under cultivate, its state broken up could be maintained, it is seen that the degree of difficulty of galactophore epithelial cell In vitro culture.
Summary of the invention
It is an object of the invention to provide the amplifying preparation process of a kind of epithelial cell, by adding IGF-in culture fluid
II, prolactin antagonist, progesterone and linoleic acid, the culture fluid configured has promotion and makees the In vitro culture of cow mammary gland epithelial cells
With.
The present invention is achieved by the following technical solutions:
The amplifying preparation process of a kind of epithelial cell, is carried out in accordance with the following steps:
Step (1), configuration epithelial cell culture fluid；
Under step (2), aseptic condition, cow mammary gland piece of tissue is washed and shredded；
After step (3), the cow mammary gland piece of tissue addition trypsin solution that will shred in described step (2) digest certain time,
Terminate digestion with the epithelial cell culture fluid of configuration in described step (1), obtain cow mammary gland tissue suspension；
Step (4), after centrifugal for the cow mammary gland tissue suspension obtained in described step (3), abandon supernatant, obtain breast
Glandular epithelium group；
Step (5), take the galactophore epithelial cell group obtained in described step (4) add in described step (1) prepare upper
Resuspended rear filtration in chrotoplast culture fluid, the culture fluid after filtering is with 1 × 105Individual/mL is inoculated in 10cm × 10cm culture dish
In, it is placed in 37 DEG C, 5%CO2Incubator is cultivated；
Suck culture fluid after step (6), original cuiture 60-72h, with hank, s liquid clean cell, add trypsin-
EDTA solution digestion 5-10min, adds the epithelial cell culture fluid of configuration in described step (1) and terminates digestion, cell suspension from
With 2 × 10 after the heart5Individual/mL is inoculated in 10cm × 10cm culture dish, is placed in 37 DEG C, 5%CO2Incubator carries out Secondary Culture.
Further, described in step (1), epithelial cell culture fluid includes basic culture solution and hyclone 10%-
20%, somatomedin 10-20 μ g/mL, kanamycin 50 μ g/mL, linoleic acid 120-180 μm ol/L.
Further, the one during described basic culture solution is DMEM culture fluid or DMEM/F12 culture fluid.
Further, described DMEM/F12 culture fluid is DMEM culture fluid and Ham, and s F12 culture fluid mixes by weight 1:1
Close.
Further, one or both mixing during described somatomedin is IGF-II, prolactin antagonist, progesterone.
Further, in described step (2), cow mammary gland piece of tissue shreds to body after phosphate buffer solution washs 3-5 time
Amass as 0.05-0.1cm3。
Further, described in step (3), trypsin solution mass fraction is 0.25%, and digestion time is 20-40min.
Further, in described step (4), centrifugation rate is 800-1500r/min, and centrifugation time is 10-30min；Described
In step (6), centrifugation rate is 1000r/min, and centrifugation time is 5-10min.
Further, described in step (6), in trypsin-EDTA solutions, trypsinase concentration is 2.5g/L, EDTA body
Fraction is 0.02%..
The method have the advantages that
A kind of epithelial cell culture fluid proposed in the present invention, adds IGF-II, prolactin antagonist, progesterone and Asia in culture fluid
Oleic acid, IGF-II tires out somatomedin as a kind of insulin, and it can stimulate beta lactoglobulin and beta-casein with prolactin antagonist
Expressing, progesterone is as the principal component of progestogen, and its major function is on the basis of estrogen action, acts on life further
Growing, promote that mammary gland tissue is reached maturity, maintain gestation and lactogenic, linoleic acid can be with inducing mammary epithelium as fatty acid
The biological activity of cell, the culture fluid configured has facilitation to the In vitro culture of cow mammary gland epithelial cells.
Certainly, the arbitrary product implementing the present invention it is not absolutely required to reach all the above advantage simultaneously.
Detailed description of the invention
Technical scheme in the embodiment of the present invention is clearly and completely described, it is clear that described embodiment is only
The a part of embodiment of the present invention rather than whole embodiments.Based on the embodiment in the present invention, those of ordinary skill in the art
The all other embodiments obtained under not making creative work premise, broadly fall into the scope of protection of the invention.
The present invention is the amplifying preparation process of a kind of epithelial cell, and the method specifically comprises the following steps that
Embodiment 1
Step (1), in DMEM culture fluid add hyclone 10%, IGF-II 20 μ g/mL, kanamycin 50 μ g/mL,
Linoleic acid 120 μm ol/L, is configured to galactophore epithelial cell culture fluid；
Under step (2), aseptic condition, cow mammary gland piece of tissue is washed 5 times through 0.1mol/L phosphate buffer solution, will wash
Cow mammary gland piece of tissue after washing shred to volume be 0.05cm3；
Step (3), the cow mammary gland piece of tissue that shreds in step (2) is added in the trypsin solution of 0.25%, and be positioned over
Digesting 40min in 37 DEG C of water bath with thermostatic control shaking tables, solid-to-liquid ratio is 1:5, obtains postdigestive cow mammary gland tissue suspension；
In postdigestive cow mammary gland tissue suspension, add the culture fluid obtained in 3mL step (1) terminate digestion；
Step (4), the cow mammary gland tissue suspension obtained in step (3) is centrifuged 10min with 1500r/min, abandons
Clear liquid, obtains galactophore epithelial cell group；
Step (5), take the galactophore epithelial cell group obtained in step (4) add in step (1) prepare culture fluid in weight
Filtering with 70 μm sterile nylon filters after Xuan, after filtering, the culture fluid containing galactophore epithelial cell group is with 1 × 105Individual/mL
It is inoculated in 10cm × 10cm culture dish, is placed in 37 DEG C, 5%CO2Incubator is cultivated；
Suck culture fluid after step (6), original cuiture 72h, clean cell 2 times with hank, s liquid, add 1mL pancreas in 37 DEG C
Protease-EDTA solution digestion 5min, in trypsin-EDTA solutions, trypsin 2.5g/L, EDTA volume fraction is
0.02%, add the culture fluid prepared in step (1) and terminate digestion；
Cell suspension is centrifuged 10min in 1000r/min, abandons supernatant, will centrifugal after galactophore epithelial cell group with 1 ×
105Individual/mL is inoculated in 10cm × 10cm culture dish, is placed in 37 DEG C, 5%CO2Incubator carries out Secondary Culture.
Embodiment 2
Step (1), in DMEM/F12 culture fluid add hyclone 20%, prolactin antagonist 10 μ g/mL, kanamycin 50 μ
G/mL, linoleic acid 180 μm ol/L, it is configured to galactophore epithelial cell culture fluid；
Under step (2), aseptic condition, cow mammary gland piece of tissue is washed 3 times through 0.1mol/L phosphate buffer solution, will wash
Cow mammary gland piece of tissue after washing shred to volume be 0.1cm3；
Step (3), the cow mammary gland piece of tissue that shreds in step (2) is added in the trypsin solution of 0.25%, and be positioned over
Digesting 20min in 37 DEG C of water bath with thermostatic control shaking tables, solid-to-liquid ratio is 1:5, obtains postdigestive cow mammary gland tissue suspension；
In postdigestive cow mammary gland tissue suspension, add the culture fluid obtained in 5mL step (1) terminate digestion；
Step (4), the cow mammary gland tissue suspension obtained in step (3) is centrifuged 30min with 800r/min, abandons supernatant
Liquid, obtains galactophore epithelial cell group；
Step (5), take the galactophore epithelial cell group obtained in step (4) add in step (1) prepare culture fluid in weight
Filtering with 70 μm sterile nylon filters after Xuan, after filtering, the culture fluid containing galactophore epithelial cell group is with 1 × 105Individual/mL
It is inoculated in 10cm × 10cm culture dish, is placed in 37 DEG C, 5%CO2Incubator is cultivated；
Suck culture fluid after step (6), original cuiture 60h, clean cell 3 times with hank, s liquid, add 0.5mL in 37 DEG C
Trypsin-EDTA solutions digestion 10min, in trypsin-EDTA solutions, trypsin 2.5g/L, EDTA volume fraction is
0.02%, add the culture fluid prepared in step (1) and terminate digestion；
Cell suspension is centrifuged 5min in 1000r/min, abandons supernatant, and the galactophore epithelial cell group after being centrifuged is with 1 × 105
Individual/mL is inoculated in 10cm × 10cm culture dish, is placed in 37 DEG C, 5%CO2Incubator carries out Secondary Culture.
Embodiment 3
Step (1), in DMEM/F12 culture fluid add hyclone 15%, prolactin antagonist 7.5 μ g/mL, progesterone 7.5 μ g/
ML, kanamycin 50 μ g/mL, linoleic acid 150 μm ol/L, it is configured to galactophore epithelial cell culture fluid；
Under step (2), aseptic condition, cow mammary gland piece of tissue is washed 4 times through 0.1mol/L phosphate buffer solution, will wash
Cow mammary gland piece of tissue after washing shred to volume be 0.75cm3；
Step (3), the cow mammary gland piece of tissue that shreds in step (2) is added in the trypsin solution of 0.25%, and be positioned over
Digesting 30min in 37 DEG C of water bath with thermostatic control shaking tables, solid-to-liquid ratio is 1:5, obtains postdigestive cow mammary gland tissue suspension；
In postdigestive cow mammary gland tissue suspension, add the culture fluid obtained in 4mL step (1) terminate digestion；
Step (4), the cow mammary gland tissue suspension obtained in step (3) is centrifuged 20min with 1200r/min, abandons
Clear liquid, obtains galactophore epithelial cell group；
Step (5), take the galactophore epithelial cell group obtained in step (4) add in step (1) prepare culture fluid in weight
Filtering with 70 μm sterile nylon filters after Xuan, after filtering, the culture fluid containing galactophore epithelial cell group is with 1 × 105Individual/mL
It is inoculated in 10cm × 10cm culture dish, is placed in 37 DEG C, 5%CO2Incubator is cultivated；
Suck culture fluid after step (6), original cuiture 66h, clean cell 3 times with hank, s liquid, in 37 DEG C of additions
0.75mL trypsin-EDTA solutions digestion 7min, trypsin 2.5g/L, EDTA volume integral in trypsin-EDTA solutions
Number is 0.02%, adds the culture fluid prepared in step (1) and terminates digestion；
Cell suspension is centrifuged 7min in 1000r/min, abandons supernatant, and the galactophore epithelial cell group after being centrifuged is with 1 × 105
Individual/mL is inoculated in 10cm × 10cm culture dish, is placed in 37 DEG C, 5%CO2Incubator carries out Secondary Culture.
A kind of galactophore epithelial cell culture fluid proposed in the present invention, adds IGF-II, prolactin antagonist, progesterone in culture fluid
And linoleic acid, IGF-II tires out somatomedin as a kind of insulin, and it can stimulate beta lactoglobulin and β-cheese egg with prolactin antagonist
White expression, progesterone is as the principal component of progestogen, and its major function is on the basis of estrogen action, acts on further
In reproductive tract, promoting that mammary gland tissue is reached maturity, maintain gestation and lactogenic, linoleic acid can be with inducing mammary as fatty acid
The biological activity of epithelial cell, the culture fluid configured has facilitation to the In vitro culture of cow mammary gland epithelial cells.
Above content is only citing made for the present invention and explanation, and affiliated those skilled in the art are to being retouched
The specific embodiment stated makes various amendment or supplements or use similar mode to substitute, without departing from inventing or super
More scope defined in the claims, all should belong to protection scope of the present invention.
Claims (8)
1. the amplifying preparation process of an epithelial cell, it is characterised in that carry out in accordance with the following steps:
Step (1), configuration epithelial cell culture fluid；
Under step (2), aseptic condition, cow mammary gland piece of tissue is washed and shredded；
After step (3), the cow mammary gland piece of tissue addition trypsin solution that will shred in described step (2) digest certain time, use institute
State the epithelial cell culture fluid of configuration in step (1) and terminate digestion, obtain cow mammary gland tissue suspension；
Step (4), after centrifugal for the cow mammary gland tissue suspension obtained in described step (3), abandon supernatant, obtain on mammary gland
Chrotoplast group；
Step (5), take the galactophore epithelial cell group obtained in described step (4) add in described step (1) prepare epithelium thin
Resuspended rear filtration in born of the same parents' culture fluid, the culture fluid after filtering is with 1 × 105Individual/mL is inoculated in 10cm × 10cm culture dish, puts
In 37 DEG C, 5%CO2Incubator is cultivated；
Suck culture fluid after step (6), original cuiture 60-72h, clean cell with hank's liquid, add trypsin-EDTA molten
Liquid digestion 5-10min, adds the epithelial cell culture fluid of configuration in described step (1) and terminates digestion, with 2 after cell suspension is centrifugal
×105Individual/mL is inoculated in 10cm × 10cm culture dish, is placed in 37 DEG C, 5%CO2Incubator carries out Secondary Culture.
The amplifying preparation process of a kind of epithelial cell the most according to claim 1, it is characterised in that: described in step (1)
Epithelial cell culture fluid includes basic culture solution and hyclone 10%-20%, somatomedin 10-20 μ g/mL, blocks that mould
Element 50 μ g/mL, linoleic acid 120-180 μm ol/L.
The amplifying preparation process of a kind of epithelial cell the most according to claim 2, it is characterised in that: described basic culture solution
For the one in DMEM culture fluid or DMEM/F12 culture fluid, described DMEM/F12 culture fluid is DMEM culture fluid and Ham's
F12 culture fluid mixes by weight 1:1.
The amplifying preparation process of a kind of epithelial cell the most according to claim 2, it is characterised in that: described somatomedin
For one or both mixing in IGF-II, prolactin antagonist, progesterone.
The amplifying preparation process of a kind of epithelial cell the most according to claim 1, it is characterised in that: in described step (2)
Cow mammary gland piece of tissue through phosphate buffer solution wash shred after 3-5 time to volume be 0.05-0.1cm3。
The amplifying preparation process of a kind of epithelial cell the most according to claim 1, it is characterised in that: described in step (3)
Trypsin solution mass fraction is 0.25%, and digestion time is 20-40min.
The amplifying preparation process of a kind of epithelial cell the most according to claim 1, it is characterised in that: in described step (4)
Centrifugation rate is 800-1500r/min, and centrifugation time is 10-30min；In described step (6), centrifugation rate is 1000r/min,
Centrifugation time is 5-10min.
The amplifying preparation process of a kind of epithelial cell the most according to claim 1, it is characterised in that: described in step (6)
In trypsin-EDTA solutions trypsinase concentration be 2.5g/L, EDTA volume fraction be 0.02%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610569366.XA CN106190952A (en) | 2016-07-19 | 2016-07-19 | A kind of amplifying preparation process of epithelial cell |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610569366.XA CN106190952A (en) | 2016-07-19 | 2016-07-19 | A kind of amplifying preparation process of epithelial cell |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106190952A true CN106190952A (en) | 2016-12-07 |
Family
ID=57493386
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610569366.XA Pending CN106190952A (en) | 2016-07-19 | 2016-07-19 | A kind of amplifying preparation process of epithelial cell |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106190952A (en) |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101407784A (en) * | 2008-11-24 | 2009-04-15 | 浙江大学 | Separation and purification method, as well as preservation method for ruminant galactophore epithelial cell |
| CN101857852A (en) * | 2010-05-31 | 2010-10-13 | 东北农业大学 | Method for building dairy cattle breast acinus lactation model in vitro |
| CN102660494A (en) * | 2012-04-17 | 2012-09-12 | 安徽农业大学 | Method for building a goat mammary epithetical cell line |
| CN103103159A (en) * | 2013-02-18 | 2013-05-15 | 天津农学院 | Cow mammary gland epithelial cell nutrient solution |
| CN103289950A (en) * | 2013-06-28 | 2013-09-11 | 浙江大学 | Methods for in vitro separation, culture, preservation and recovery of milk cow omasum epithelial cells |
| CN104195100A (en) * | 2014-08-29 | 2014-12-10 | 山东省农业科学院畜牧兽医研究所 | In-vitro culture method of mammary gland epithelial cells |
| CN105316279A (en) * | 2014-07-17 | 2016-02-10 | 北京大北农科技集团股份有限公司动物医学研究中心 | Method for efficiently separating and purifying mammary epithelial cells |
| CN105543164A (en) * | 2016-02-29 | 2016-05-04 | 西北农林科技大学 | Primary isolated culture method for dairy cow mammary epithelial cells |
| CN105754933A (en) * | 2016-04-21 | 2016-07-13 | 中国农业科学院兰州畜牧与兽药研究所 | Cell in-vitro culture method for primary culture of bovine mammary epithelial cells |
-
2016
- 2016-07-19 CN CN201610569366.XA patent/CN106190952A/en active Pending
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101407784A (en) * | 2008-11-24 | 2009-04-15 | 浙江大学 | Separation and purification method, as well as preservation method for ruminant galactophore epithelial cell |
| CN101857852A (en) * | 2010-05-31 | 2010-10-13 | 东北农业大学 | Method for building dairy cattle breast acinus lactation model in vitro |
| CN102660494A (en) * | 2012-04-17 | 2012-09-12 | 安徽农业大学 | Method for building a goat mammary epithetical cell line |
| CN103103159A (en) * | 2013-02-18 | 2013-05-15 | 天津农学院 | Cow mammary gland epithelial cell nutrient solution |
| CN103289950A (en) * | 2013-06-28 | 2013-09-11 | 浙江大学 | Methods for in vitro separation, culture, preservation and recovery of milk cow omasum epithelial cells |
| CN105316279A (en) * | 2014-07-17 | 2016-02-10 | 北京大北农科技集团股份有限公司动物医学研究中心 | Method for efficiently separating and purifying mammary epithelial cells |
| CN104195100A (en) * | 2014-08-29 | 2014-12-10 | 山东省农业科学院畜牧兽医研究所 | In-vitro culture method of mammary gland epithelial cells |
| CN105543164A (en) * | 2016-02-29 | 2016-05-04 | 西北农林科技大学 | Primary isolated culture method for dairy cow mammary epithelial cells |
| CN105754933A (en) * | 2016-04-21 | 2016-07-13 | 中国农业科学院兰州畜牧与兽药研究所 | Cell in-vitro culture method for primary culture of bovine mammary epithelial cells |
Non-Patent Citations (3)
| Title |
|---|
| 朝格图: "奶牛乳腺上皮细胞和成纤维细胞原代培养与鉴定", 《 中国优秀硕士学位论文全文数据库 基础科学辑》 * |
| 杨文浩等: "酶消化组织块法体外培养奶牛乳腺上皮细胞", 《畜牧与兽》 * |
| 王治国: "奶牛乳腺上皮细胞的体外培养及应用", 《 中国优秀硕士学位论文全文数据库 基础科学辑》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107354129B (en) | A kind of Fibroblast cell-culture method in autologous skin source | |
| CN104762257B (en) | A kind of method preparing mescenchymal stem cell from umbilical cord | |
| CN102732586B (en) | Method for culturing mesenchymal stem cell secretin | |
| CN102660494A (en) | Method for building a goat mammary epithetical cell line | |
| CN106038598A (en) | Method for preparing human-derived stem cell secretion bioactive factor and lysate | |
| CN105969726A (en) | Method for preparing adipose-derived stem cells by means of extraction | |
| CN101407784B (en) | Separation and purification method, as well as preservation method for ruminant galactophore epithelial cell | |
| CN102146359A (en) | Method for extracting original mesenchymal stem cells from placenta and serum-free amplification | |
| CN103756952A (en) | Establishment and application method of ovary cell line of cynoglossus semilaevis | |
| CN101597592B (en) | Human corneal endothelial cell culture solution as well as preparation method and application thereof | |
| CN109294972A (en) | A kind of lamb intestinal epithelial cell extracorporeal culturing method | |
| CN104971382B (en) | The mounted artificial active mass of wound and its construction method that a kind of use serum-free and ox pituitary extract nutrient solution are built | |
| CN102925407B (en) | In-vitro culture method of triploid loach fin cells | |
| CN106190952A (en) | A kind of amplifying preparation process of epithelial cell | |
| CN104946579A (en) | Culture method of Jinhua pig mammary gland epithelial cell line | |
| CN107129966A (en) | A kind of corneal epithelial cell nutrient solution containing serum | |
| CN104195100A (en) | In-vitro culture method of mammary gland epithelial cells | |
| CN106075585B (en) | A kind of preparation method based on artificial tendon scaffold materials microstructure engineering artificial tendon graft | |
| CN104771788B (en) | A kind of organization engineering skin and its construction method based on omentum majus acellular matrix | |
| WO2022068029A1 (en) | Special culture apparatus for 3d biological tissue, and method for preparing block-shaped cultured meat | |
| CN109771697B (en) | Dermal fibroblast skin sheet and construction method and application thereof | |
| CN101861857B (en) | Preservation method of ruminant mammary gland epithelial cells | |
| CN114736854A (en) | Culture method of stem cells | |
| CN201634683U (en) | Device for separating primary cells from tissue | |
| CN102559589A (en) | Method for performing in-vitro culture to bovine preadipocytes |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20161207 |
|
| RJ01 | Rejection of invention patent application after publication |