CN106256900A - A kind of stem cell cultivating system of non-animal derived property - Google Patents
A kind of stem cell cultivating system of non-animal derived property Download PDFInfo
- Publication number
- CN106256900A CN106256900A CN201610666606.8A CN201610666606A CN106256900A CN 106256900 A CN106256900 A CN 106256900A CN 201610666606 A CN201610666606 A CN 201610666606A CN 106256900 A CN106256900 A CN 106256900A
- Authority
- CN
- China
- Prior art keywords
- stem cell
- animal derived
- cultivating system
- derived property
- cell cultivating
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0662—Stem cells
- C12N5/0663—Bone marrow mesenchymal stem cells (BM-MSC)
-
- 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
- C12N2500/00—Specific components of cell culture medium
- C12N2500/30—Organic components
- C12N2500/32—Amino acids
-
- 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
- C12N2500/44—Thiols, e.g. mercaptoethanol
-
- 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
- C12N2500/46—Amines, e.g. putrescine
-
- 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
- C12N2501/11—Epidermal growth factor [EGF]
-
- 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
- C12N2501/115—Basic fibroblast growth factor (bFGF, FGF-2)
-
- 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
- C12N2501/20—Cytokines; Chemokines
- C12N2501/23—Interleukins [IL]
- C12N2501/235—Leukemia inhibitory factor [LIF]
-
- 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
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/90—Polysaccharides
- C12N2501/91—Heparin
-
- 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
- C12N2501/998—Proteins not provided for elsewhere
-
- 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
- C12N2513/00—3D culture
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Developmental Biology & Embryology (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Cell Biology (AREA)
- Microbiology (AREA)
- Rheumatology (AREA)
- Hematology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Immunology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses the stem cell cultivating system of a kind of non-animal derived property, forming including feeder layer cells and the growth medium being made by HFF, growth medium includes that DMEM culture medium, the pla-pcl three-dimensional rack being made, serum substitute, β dredge base ethanol, LIF, non essential amino acid, Collagen type Ⅳ, insulin and antibiotic.The present invention has the following advantages and effect: by adding the feeder layer cells that HFF is made in cultivating system, avoid pollution that animal derived materials causes and uncertain composition, by adding the LIF of suppression stem cell differentiation and promoting that the insulin of stem cells hyperplasia and β dredge base ethanol, human serum or animal serum is replaced with serum substitute, the three-dimensional rack being made with pla-pcl simulates the three-dimensional environment that stem cell is residing in vivo, improve stem cell culture efficiency, reach to remove animal derived materials, cultivate composition and cultivation mechanism is clear and definite, culture efficiency is high effect.
Description
Technical field
The present invention relates to stem cell and cultivate field, particularly to the stem cell cultivating system of a kind of non-animal derived property.
Background technology
Stem cell is the pluripotent cell that a class has the of self-replication capacity.Under certain condition, it can be divided into many
Plant functioning cell.It is divided into embryonic stem cell and adult stem cell according to the stage of development residing for stem cell.Sending out according to stem cell
Educate potential to be divided three classes: myeloid-lymphoid stem cell, pluripotent stem cell and unipotent stem cell.
The Chinese patent " a kind of stem cell culture method " of Publication No. CN101880649A discloses a kind of stem cell training
Breeding method, prepares without losing the amniotic epithelial cells trophoderm that splitting ability processes；Inoculation stem cell is in amniotic epithelial cells
On trophoderm, cultivate in culture fluid.
But this stem cell culture method has the disadvantage in that amniotic epithelial cells trophoderm can comprise the unknown in secretions
The factor and composition, can comprise animal derived materials, after adding human whole serum or cord serum when cultivating simultaneously, containing unknown because of
Son likely differentiation of stem cells or death, trophoderm and human whole serum can cause animal derived materials in culture medium after adding
With uncertain composition, limit application and the research of stem cell.
Summary of the invention
It is an object of the invention to provide the stem cell cultivating system of a kind of non-animal derived property, the effect that tool composition determines.
The above-mentioned technical purpose of the present invention has the technical scheme that the dry thin of a kind of non-animal derived property
Born of the same parents' cultivating system, forms including feeder layer cells and the growth medium being made by HFF, and described growth medium includes
DMEM culture medium, the pla-pcl three-dimensional rack being made, serum substitute, β-dredge base ethanol, LIF, non-essential amino
Acid, Collagen type Ⅳ, insulin and antibiotic.
Optimize stem cell cultivating system research at stem cell scientific research field achieved with many progress, but remove animal become
The raising of impact, clear and definite, the shortening of cultivation cycle of culture medium culturing mechanism and the efficiency divided still is difficult to every taking into account.
The present invention uses the feeder layer cells that HFF is made, and makes the pollution of non-animal derived property in cultivating system, in production medium
Do not use the uncertain human serum of composition or animal serum, but use serum substitute, compared to human serum and animal serum,
Serum substitute not only definite ingredients, and with the feeder layer cells synergy in cultivating system, can effectively maintain stem cell
Self renewal and propagation, i.e. promote the propagation of stem cell on the premise of keeping stem cell dryness.
DMEM culture medium is selected to provide the various materials needed for growth for stem cell, it is ensured that nutrition supplying during stem cells hyperplasia
Give.General stem cell growth and split speed are very fast, and cell often divides once, it is necessary to synthesize substantial amounts of protein and nucleic acid,
Only make the non essential amino acid of cell self synthesis dry up, not only can not meet demand, but also the quick increasing of stem cell can be limited
Grow.By additionally adding non essential amino acid in cultivating system, improve stem cells hyperplasia speed.
Pla-pcl is the linear aliphatic adoption ester obtained by ε-caprolactone ring-opening polymerisation, has biology in cell is cultivated
The compatibility is good, toxicity is low, and the beneficially propagation of cell and growth.Propped up by the three-dimensional utilizing pla-pcl to be made
Frame, simulates the three-dimensional environment that stem cell is residing in vivo, plays the effect of similar scaffold, be attached to three during stem cell growth
Grow on dimensional scaffold and migrate.After adding Laminin lens in cultivating system, Laminin lens is attached to three-dimensional rack table
Face, containing the site combined for cell membrane on Laminin lens, plays on three-dimensional rack attached for stem cell as connecting medium
The effect.It addition, Laminin lens is in addition to adhesive attraction, promotion stem cells hyperplasia and the work of Inhibited differentiation also can be played
With.
While stem cells hyperplasia, the differentiation of suppression stem cell is a primary condition, and LIF Chinese is translated into leukemia to be pressed down
The factor processed, it is a kind of cytokine with several functions, but its most important application is to maintain embryonic stem cell etc. not
Differentiation state.During stem cells hyperplasia, LIF can suppress the differentiation of stem cell, it is to avoid the stem cell shadow after stem cell differentiation and differentiation
Ring remaining undifferentiated stem cell.
Keeping stem cell undifferentiated while, keep Stem Cell Activity and improve culture efficiency, to Optimized culture system
There is significant benefit.Growth medium adds β-thin base ethanol, the propagation of stem cell is had obvious facilitation.With
Time regulate stem cell division time, insulin like growth factor has regulation effect to the division of stem cell, but due in reality
During trampling, insulin like growth factor extracts the most difficult, and phase in insulin and insulin like growth factor not only structure
Seemingly, the most also there is concordance, replace insulin like growth factor that stem cell division is adjusted by adding insulin,
The propagation promoting stem cell is worked in coordination with β-thin base ethanol.In cultivating system, add antibiotic, prevent incubation antibacterial etc. micro-
Biological intrusion and pollution.
The further setting of the present invention is: in described production medium, described serum substitute is 10%, described β-dredge base
Concentration of alcohol is 0.1mmol/ml, and described LIF concentration is 1.5 × 103U/ml, described antibiotic be 60mg/L penicillin and
The streptomycin of 100mg/L, described insulin concentration is 20ng/ml.
By using technique scheme, insulin concentration is 20ng/ml, makes insulin can adjust stem cells hyperplasia
Joint, reduces the toxicity to stem cell.β-dredge base concentration of alcohol be 0.1mmol/ml, LIF concentration be 1.5 × 103U/ml, antibiotic
For penicillin and the streptomycin of 100mg/L of 60mg/L, both can guarantee that the growth to stem cell had biological activity, avoided again concentration
Too high and cause the damage to stem cell.
The further setting of the present invention is: described growth medium also includes the human albumin of 10g/L.
By using technique scheme, although do not add human serum or animal serum, but determining cultivating system
On the premise of composition, the propagation of stem cell can be promoted after adding human albumin, the differentiation of stem cell can be suppressed again.
The further setting of the present invention is: described growth medium also includes the progesterone of 10nmol/l.
By using technique scheme, progesterone can stimulate the propagation of cell, and can promote that stem cell is along three-dimensional rack
Migrate, improve the growth rate of stem cell further.
The further setting of the present invention is: described productive culture machine also includes the sodium selenite of 10nmol/l.
By using technique scheme, in stem cell incubation, stem cell is when metabolism tyrosine, tryptophan etc.
Can produce hydrogen peroxide, hydrogen peroxide can produce toxicity to stem cell, makes stem cell silk screen, adds sub-selenium in growth medium
Acid sodium, the selenium in sodium selenite participates in glutathion peroxidase and the mechanism of superoxide dismutase, promotes peroxide
The degraded to hydrogen peroxide of the compound enzyme, reduces the apoptosis rate of stem cell.
The further setting of the present invention is: described growth medium also includes the butanediamine of 20 μm ol/l.
By using technique scheme, butanediamine is required when being the lipid needed for cell membrane synthesizes and cell proliferation division
Water solublity lipid, add after butanediamine the propagation to stem cell and further function as facilitation.
The further setting of the present invention is: described growth medium also includes Wnt protein and Heparan sulfate.
By using technique scheme, Wnt protein is a kind of protein promoting stem cell to update, acetyl sulfate liver
Element is a kind of carbohydrate, and Wnt protein can be assisted after addition to transmit signal to stem cell, and stem cell will not be divided in suppression
Other cells.
The further setting of the present invention is: described growth medium also includes the EGF and the FGF-2 of 20 μ g/L of 20 μ g/L.
By using technique scheme, EGF and FGF-2 synergism, promote the propagation of stem cell, improve training further
The efficiency supported.
The further setting of the present invention is: described DMEM culture medium is high glycoform DMEM culture medium.
By using technique scheme, the fast breeding of stem cell is improved sufficient sugar by the DMEM culture medium of high glycoform
Point, the differentiation of stem cell can be suppressed simultaneously.
In sum, the method have the advantages that by adding the raising that HFF is made in cultivating system
Confluent monolayer cells, avoids pollution that animal derived materials causes and uncertain composition, by adding LIF and the rush of suppression stem cell differentiation
Enter insulin and the β-thin base ethanol of stem cells hyperplasia, by adding Wnt protein and Heparan sulfate, promote dry further
The propagation of cell, replaces human serum or animal serum with serum substitute, specifies the composition in cultivating system, use DMEM culture medium
The propagation being stem cell with non essential amino acid provides material, and the three-dimensional rack being made with pla-pcl simulates stem cell
Three-dimensional environment residing in vivo, improves stem cell culture efficiency, reaches to remove animal derived materials, cultivate composition and culture machine
The effect that system is clearly, culture efficiency is high.
Detailed description of the invention
Specific embodiment is only explanation of the invention, and it is not limitation of the present invention, those skilled in the art
As required the present embodiment can be made after reading this specification and there is no the amendment of creative contribution, but as long as at this
All protected by Patent Law in bright right.
Embodiment 1: the stem cell cultivating system of a kind of non-animal derived property, is purchased HFF, i.e. human foreskin fibroblast, will
It processes through mitotic block, after being fabricated to feeder layer cells, is inoculated in Tissue Culture Dish, then trains with high glycoform DMEM
Based on supporting base, each proportions in table 1 below goes out production medium, the three-dimensional framework being made by pla-pcl, uses
Feeder layer cells, three-dimensional rack, growth medium form the stem cell cultivating system of non-animal derived property.In prior art disclosed
The dimensional culture system being made with pla-pcl.Stem cell selects the people's bone marrow mesenchymal stem cell being purchased, and is not related to appoint
What ethics.The stem cell cultivating system that people's bone marrow mesenchymal stem cell is inoculated in during use above-mentioned non-animal derived property is cultivated
?.
Cell proliferation test: be purchased BrdU cell proliferation Assay test kit, from Chemicon
International.Choose addition BrdU labelling in stem cell cultivating system, after hatching 4 hours, draw to get rid of and train completely
After supporting base, add fixative greenhouse and hatch 30 minutes, make cell DNA degeneration, the washing provided with test kit after isolating cell
Liquid washed cell 5 minutes, after washing 3 times continuously, joins the anti-BrdU incubated at room that test kit provides after isolating cell
1 hour, add Goat anti-mouse IgG-POD, incubated at room 30 minutes after again separating cell, be eventually adding test kit
The acid reaction stop buffer provided, reads absorbance by microplate reader at wavelength 450nm, and absorbance is the highest, represents dry thin
The BrdU concentration mixed in born of the same parents is the highest, and reflection stem cells hyperplasia is the most active.And list testing result in Table 1.
Cell morphological characteristic detection test: the form of people's bone marrow mesenchymal stem cell is elongated fusiformis, the cell after differentiation
Form is then roomy pancake, and therefore the cellular morphology of detection people's bone marrow mesenchymal stem cell i.e. can detect whether differentiation.With purchasing
From the cellular morphology of inverted microscope observer's bone marrow mesenchymal stem cell of Nikon, and list testing result in Table 1.
Embodiment 2: the stem cell cultivating system of a kind of non-animal derived property, is with the difference of embodiment 1, growth training
Support base to prepare by each component in table 1 below, and carry out cell proliferation test and cell morphological characteristic detection test, and
Testing result listed by table 1.
Embodiment 3: the stem cell cultivating system of a kind of non-animal derived property, is with the difference of embodiment 1, growth training
Support base to prepare by each component in table 1 below, and carry out cell proliferation test and cell morphological characteristic detection test, and
Testing result listed by table 1.
Embodiment 4: the stem cell cultivating system of a kind of non-animal derived property, is with the difference of embodiment 1, growth training
Support base to prepare by each component in table 1 below, and carry out cell proliferation test and cell morphological characteristic detection test, and
Testing result listed by table 1.
Embodiment 5: the stem cell cultivating system of a kind of non-animal derived property, is with the difference of embodiment 1, growth training
Support base to prepare by each component in table 1 below, and carry out cell proliferation test and cell morphological characteristic detection test, and
Testing result listed by table 1.
Embodiment 6: the stem cell cultivating system of a kind of non-animal derived property, is with the difference of embodiment 1, growth training
Support base to prepare by each component in table 1 below, and carry out cell proliferation test and cell morphological characteristic detection test, and
Testing result listed by table 1.
Embodiment 7: the stem cell cultivating system of a kind of non-animal derived property, is with the difference of embodiment 1, growth training
Support base to prepare by each component in table 1 below, and carry out cell proliferation test and cell morphological characteristic detection test, and
Testing result listed by table 1.
Embodiment 8: the stem cell cultivating system of a kind of non-animal derived property, is with the difference of embodiment 1, growth training
Support base to prepare by each component in table 1 below, and carry out cell proliferation test and cell morphological characteristic detection test, and
Testing result listed by table 1.
Comparative example 1: the stem cell cultivating system of a kind of non-animal derived property, is with the difference of embodiment 1, growth training
Support base to prepare by each component in table 1 below, and carry out cell proliferation test and cell morphological characteristic detection test, and
Testing result listed by table 1.
Contrast 2: the stem cell cultivating system of a kind of non-animal derived property, is with the difference of embodiment 1, grown cultures
Base is prepared by each component in table 1 below, and carries out cell proliferation test and cell morphological characteristic detection test, and at table
Testing result is listed in 1.Comparative example 3: the stem cell cultivating system of a kind of non-animal derived property, exists with the difference of embodiment 1
In, growth medium is prepared by each component in table 1 below, and carries out cell proliferation test and cell morphological characteristic detection
Test, and list testing result in Table 1.
Table 1
Claims (9)
1. the stem cell cultivating system of a non-animal derived property, it is characterised in that: include the feeder layer cells being made by HFF
Forming with growth medium, described growth medium includes DMEM culture medium, the pla-pcl three-dimensional rack being made, blood
Clear substitute, β-dredge base ethanol, LIF, non essential amino acid, Collagen type Ⅳ, insulin and antibiotic.
The stem cell cultivating system of a kind of non-animal derived property the most according to claim 1, it is characterised in that: described production is trained
Supporting described serum substitute in base is 10%, and described β-dredge base concentration of alcohol is 0.1mmol/ml, described LIF concentration is 1.5 ×
103U/ml, described antibiotic is penicillin and the streptomycin of 100mg/L of 60mg/L, and described insulin concentration is 20ng/ml.
The stem cell cultivating system of a kind of non-animal derived property the most according to claim 2, it is characterised in that: described growth training
Foster base also includes the human albumin of 10g/L.
The stem cell cultivating system of a kind of non-animal derived property the most according to claim 3, it is characterised in that: described growth training
Foster base also includes the progesterone of 10nmol/l.
The stem cell cultivating system of a kind of non-animal derived property the most according to claim 4, it is characterised in that: described production is trained
Foster machine also includes the sodium selenite of 10nmol/l.
The stem cell cultivating system of a kind of non-animal derived property the most according to claim 5, it is characterised in that: described growth training
Foster base also includes the butanediamine of 20 μm ol/l.
The stem cell cultivating system of a kind of non-animal derived property the most according to claim 6, it is characterised in that: described growth training
Foster base also includes Wnt protein and Heparan sulfate.
The stem cell cultivating system of a kind of non-animal derived property the most according to claim 7, it is characterised in that: described growth training
Foster base also includes the EGF and the FGF-2 of 20 μ g/L of 20 μ g/L.
The stem cell cultivating system of a kind of non-animal derived property the most according to claim 8, it is characterised in that: described DMEM trains
Foster base is high glycoform DMEM culture medium.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610666606.8A CN106256900A (en) | 2016-08-12 | 2016-08-12 | A kind of stem cell cultivating system of non-animal derived property |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610666606.8A CN106256900A (en) | 2016-08-12 | 2016-08-12 | A kind of stem cell cultivating system of non-animal derived property |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106256900A true CN106256900A (en) | 2016-12-28 |
Family
ID=57713673
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610666606.8A Pending CN106256900A (en) | 2016-08-12 | 2016-08-12 | A kind of stem cell cultivating system of non-animal derived property |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106256900A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113481147A (en) * | 2021-06-10 | 2021-10-08 | 艾可泰科生物科技(江苏)有限公司 | Method for constructing pancreatic stem cell line from human pancreatic islets and differentiating pancreatic islet cells |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1626651A (en) * | 2003-12-10 | 2005-06-15 | 财团法人工业技术研究院 | System for culturing human embryonic stem cell and culturing method |
| CN101415818A (en) * | 2006-04-04 | 2009-04-22 | 3M创新有限公司 | Flat microfibers as matrices for cell growth |
| CN103555661A (en) * | 2013-10-25 | 2014-02-05 | 中国科学院动物研究所 | Culture method free of multipotential stem cell without serum and feeder layer |
| CN104140951A (en) * | 2014-08-06 | 2014-11-12 | 深圳市科晖瑞生物医药有限公司 | Method for developing and cultivating human induced pluripotent stem cells |
| CN105087479A (en) * | 2015-08-21 | 2015-11-25 | 深圳爱生再生医学科技有限公司 | Stem cell serum-free culture medium and stem cell culture method |
| CN105112365A (en) * | 2015-08-18 | 2015-12-02 | 广州暨南生物医药研究开发基地有限公司 | Serum-free medium for human umbilical cord mesenchymal stem cells and preparation method thereof |
-
2016
- 2016-08-12 CN CN201610666606.8A patent/CN106256900A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1626651A (en) * | 2003-12-10 | 2005-06-15 | 财团法人工业技术研究院 | System for culturing human embryonic stem cell and culturing method |
| CN101415818A (en) * | 2006-04-04 | 2009-04-22 | 3M创新有限公司 | Flat microfibers as matrices for cell growth |
| CN103555661A (en) * | 2013-10-25 | 2014-02-05 | 中国科学院动物研究所 | Culture method free of multipotential stem cell without serum and feeder layer |
| CN104140951A (en) * | 2014-08-06 | 2014-11-12 | 深圳市科晖瑞生物医药有限公司 | Method for developing and cultivating human induced pluripotent stem cells |
| CN105112365A (en) * | 2015-08-18 | 2015-12-02 | 广州暨南生物医药研究开发基地有限公司 | Serum-free medium for human umbilical cord mesenchymal stem cells and preparation method thereof |
| CN105087479A (en) * | 2015-08-21 | 2015-11-25 | 深圳爱生再生医学科技有限公司 | Stem cell serum-free culture medium and stem cell culture method |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113481147A (en) * | 2021-06-10 | 2021-10-08 | 艾可泰科生物科技(江苏)有限公司 | Method for constructing pancreatic stem cell line from human pancreatic islets and differentiating pancreatic islet cells |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Guan et al. | Trends and ideas in technology, regulation and public acceptance of cultured meat | |
| CN103060264B (en) | Stem cell culture medium and application thereof and stem cell cultivation method | |
| CN103243070B (en) | Stem cell medium and application thereof | |
| CN103857789B (en) | Prepare mescenchymal stem cell basal medium and utilize the method that mescenchymal stem cell basal medium prepares cell therapy product and the differentiation product obtained by this culture medium | |
| WO2015180636A1 (en) | Specific medium for long-term maintenance and proliferation subculture of human hepatocytes and culture method | |
| CN102186969A (en) | Methods for preparing human skin substitutes from human pluripotent stem cells | |
| KR102445537B1 (en) | System and method for 3d cell culture and use thereof | |
| Serban et al. | Effects of extracellular matrix analogues on primary human fibroblast behavior | |
| CN104988110A (en) | Method for transforming umbilical cord mesenchymal stem cells into islet cells | |
| Li et al. | Construction of bioengineered hepatic tissue derived from human umbilical cord mesenchymal stem cells via aggregation culture in porcine decellularized liver scaffolds | |
| US20190264179A1 (en) | Serum-free medium inducing differentiation of umbilical cord mesenchymal stem cell into insulin-secretion-like cell and preparation method and use thereof | |
| CN106459913A (en) | Improved stem cell composition | |
| Zhang et al. | Comparison of beneficial factors for corneal wound-healing of rat mesenchymal stem cells and corneal limbal stem cells on the xenogeneic acellular corneal matrix in vitro | |
| CN103497892B (en) | A kind of cell cultures base material and its preparation method and application | |
| RU2019134339A (en) | ISOLATION OF CELLS FROM HATCHED EGGS OF REPTILES FOR USE TO OBTAIN BIOLOGICAL DERMAS AND SKIN FOR SKIN PRODUCTION | |
| Yuen Jr et al. | Perspectives on scaling production of adipose tissue for food applications | |
| Bhatia et al. | Introduction to animal tissue culture science | |
| David et al. | Co-culture approaches for cultivated meat production | |
| Zhang et al. | Generation of functional fat organoid from rat superficial fascia | |
| CN107312744B (en) | Serum-containing oral mucosa epithelial cell culture solution | |
| CN104988111A (en) | Inducing liquid for converting UC-MSC into islet cells and application thereof | |
| CN108753710A (en) | A kind of Serum-free complete medium and its application | |
| CN106256900A (en) | A kind of stem cell cultivating system of non-animal derived property | |
| CN106244522A (en) | A kind of stem cell cultivating system and cultural method thereof | |
| CN105886462A (en) | Composition ADSCs for ADSCs culture and ADSCs culture method |
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 | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20161228 |