CN106011053A - Novel dental pulp stem cell culturing medium - Google Patents
Novel dental pulp stem cell culturing medium Download PDFInfo
- Publication number
- CN106011053A CN106011053A CN201610475763.0A CN201610475763A CN106011053A CN 106011053 A CN106011053 A CN 106011053A CN 201610475763 A CN201610475763 A CN 201610475763A CN 106011053 A CN106011053 A CN 106011053A
- Authority
- CN
- China
- Prior art keywords
- stem cell
- dental pulp
- pulp stem
- culturing medium
- culture medium
- 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
- C12N5/0664—Dental pulp stem cells, Dental follicle stem cells
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
- C12N2501/15—Transforming growth factor beta (TGF-β)
Abstract
The invention discloses a novel dental pulp stem cell culturing medium. Every L of the novel dental pulp stem cell culturing medium is prepared from 100-150 ml of fetal calf serum, 10-15 ml of penicillin-streptomycin antibodies, 1-5 ng of TGF-beta, 1-5 mM of L-glutamine and the balance 1640 culturing media. A preparing method includes the steps that the TGF-beta, the L-glutamine and the penicillin-streptomycin antibodies are added into 1/3 of the whole 1640 culturing medium, the mixture is stirred and dissolved, the left 1640 culturing medium is added, and the volume is fixed to be 1 L, and the product is stirred to be even; the prepared solution is filtered with a 0.22-um filtering film, the fetal calf serum is added under aseptic conditions, and the novel dental pulp stem cell culturing medium is obtained. According to the culturing media, the basic culturing medium is 1640 and contains inorganic constituents required when deciduous-tooth dental pulp matrix cells are differentiated into odontoblast; the culturing medium contains the fetal calf serum, the nutrient substances required by cell growth are provided, and cell adherence growth can be promoted; the glutamine and the TGF-beta can promote cell propagation; the penicillin-streptomycin antibodies have the antibacterial effect.
Description
Technical field
The present invention relates to stem cell and cultivate field, particularly relate to novel dental pulp stem cell media.
Background technology
Stem cell is the pluripotent cell that a class has the of self-replication capacity, is the cell of original and non-specialization.Necessarily
Under the conditions of, it can be divided into several functions cell, has the various histoorgans of regeneration and the potential function of human body.Stem cell is deposited
In all multicellular tissues, can via mitosis with split into multiple specialized cell, and can be utilized oneself
I more newly arrives and provides more stem cell.The source of stem cell has a lot, including bone marrow, umbilical cord, Cord blood, tooth and fat.
Within 2000, being found that dental pulp stem cell first, Gronthos etc. finds to there is tooth in vitro deciduous teeth and wisdom teeth
Marrow stem cell.This is that a class has high proliferation ability, highly self-renewal capacity, the cell of multinomial differentiation capability.Study
This cell is utilized successfully to build dental pulp, dentin and dental tissue.The stem cell that dental pulp stem cell is tissue-derived with other
Biological characteristics has a lot of similarity.The acquisition of deciduous teeth is the most painless, and everyone is longer
Be 20 deciduous teeth, deciduous teeth i.e. started eruption at about half years old, within about 2 years old half, all sprouted, and between 6-12 year, deciduous teeth come off successively, because of
This obtains mescenchymal stem cell from deciduous teeth dental pulp will be more and more universal.
2003, Miura etc. isolates from the dental pulp of the deciduous teeth that come off had high proliferative capacity and Multidirectional Differentiation ability
Stem cell-SHED.Owing to it has a biological characteristics of good mescenchymal stem cell, tissue-derived it is easy to get and hurtless measure,
It is considered as that stem cell tissue engineering carries out the seed cell that regeneration and restoration is more satisfactory.
Modern study finds that deciduous teeth mescenchymal stem cell is at treatment bone and dental tissue Regeneration and Repair, nervous system disease and flesh
The aspects such as meat atrophy, disease of immune system, skin injury, hepatic disease have broad application prospects.Therefore, exploitation is simple, fast
Speed, a large amount of deciduous teeth mescenchymal stem cell methods of safety acquisition become the task of top priority, in the cultivation of stem cell, and the selection of culture medium
Being the research on basis, suitable culture medium selects to be to do stem cell to cultivate successfully guarantee.
Summary of the invention
It is an object of the invention to provide a kind of novel dental pulp stem cell media.
It is an object of the invention to be achieved through the following technical solutions: a kind of novel dental pulp stem cell media, every 1L
Dental pulp stem cell culture medium consists of the following composition:
Hyclone 100-150ml
The dual anti-10-15ml of mycillin
TGF-β 1-5ng
L-glutaminate 1-5mM
1640 culture medium surpluses.
On the basis of such scheme preferably, every 1L dental pulp stem cell culture medium consists of the following composition:
Hyclone 120ml
The dual anti-12ml of mycillin
TGF-β 3ng
L-glutaminate 3mM
1640 culture medium surpluses.
A kind of method preparing dental pulp stem cell culture medium, comprises the following steps:
Step one, takes 1640 culture medium of total amount 1/3, and addition TGF-β, L-glutaminate, mycillin are dual anti-, stirring and dissolving,
Add remaining 1640 culture medium and be settled to 1L, stir；
Step 2, the solution 0.22um membrane filtration that step one is prepared, add hyclone under aseptic condition, to obtain final product；With
0.22um membrane filtration, can reach good degerming purpose.
Beneficial effects of the present invention: the culture medium in the present invention, basal medium is 1640, and it contains deciduous teeth pulp matrix
Cell inorganic constituents required for the differentiation of odontoblast direction；Containing hyclone in culture medium, it is provided that cell grows
Desired nutritional material also can promote that cell attachment grows；Glutamine and TGF-β can strengthen the propagation of cell；Mycillin
Dual anti-play antibacterial action.
Detailed description of the invention
Below in conjunction with specific embodiment, the invention will be further described:
Embodiment 1
Preparation 1L novel dental pulp stem cell media:
Take hyclone 100ml, the dual anti-10ml of mycillin, TGF-β (sigma) 5ng, L-glutaminate (sigma) 5mM,
1640 culture medium 1L；
Take 1640 culture medium of 0.3L, add TGF-β (sigma), M L-glutaminate (sigma), mycillin dual anti-
(sigma), stirring and dissolving, add 1640 culture medium to 1L constant volume, stir, degerming with 0.22um membrane filtration, aseptic bar
Add hyclone (gibco) under part, preserve in prepared solution sterile subpackage to sterilized receiving flask.
Embodiment 2
Preparation 1L novel dental pulp stem cell media:
Take hyclone 120ml, the dual anti-12ml of mycillin, TGF-β (sigma) 3ng, L-glutaminate (sigma) 3mM,
1640 culture medium 1L；
Take 1640 culture medium of 0.3L, add TGF-β (sigma), M L-glutaminate (sigma), mycillin dual anti-
(sigma), stirring and dissolving, add 1640 culture medium to 1L constant volume, stir, degerming with 0.22um membrane filtration, aseptic bar
Add hyclone (gibco) under part, preserve in prepared solution sterile subpackage to sterilized receiving flask.
Embodiment 3
Preparation 1L novel dental pulp stem cell media:
Take hyclone 150ml, the dual anti-15ml of mycillin, TGF-β (sigma) 1ng, L-glutaminate (sigma) 1mM,
1640 culture medium 1L；
Take 1640 culture medium of 0.3L, add TGF-β (sigma), M L-glutaminate (sigma), mycillin dual anti-
(sigma), stirring and dissolving, add 1640 culture medium to 1L constant volume, stir, degerming with 0.22um membrane filtration, aseptic bar
Add hyclone (gibco) under part, preserve in prepared solution sterile subpackage to sterilized receiving flask.
Claims (3)
1. a novel dental pulp stem cell media, it is characterised in that by following component group in every 1L dental pulp stem cell culture medium
Become:
Hyclone 100-150ml
The dual anti-10-15ml of mycillin
TGF-β 1-5ng
L-glutaminate 1-5mM
1640 culture medium surpluses.
Novel dental pulp stem cell media the most according to claim 1, it is characterised in that every 1L dental pulp stem cell is cultivated
Base consists of the following composition:
Hyclone 120ml
The dual anti-12ml of mycillin
TGF-β 3ng
L-glutaminate 3mM
1640 culture medium surpluses.
3. the method preparing dental pulp stem cell culture medium, it is characterised in that comprise the following steps:
Step one, takes 1640 culture medium of total amount 1/3, and addition TGF-β, L-glutaminate, mycillin are dual anti-, stirring and dissolving,
Add remaining 1640 culture medium and be settled to 1L, stir；
Step 2, the solution 0.22um membrane filtration that step one is prepared, add hyclone under aseptic condition, to obtain final product.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610475763.0A CN106011053A (en) | 2016-06-27 | 2016-06-27 | Novel dental pulp stem cell culturing medium |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610475763.0A CN106011053A (en) | 2016-06-27 | 2016-06-27 | Novel dental pulp stem cell culturing medium |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106011053A true CN106011053A (en) | 2016-10-12 |
Family
ID=57083607
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610475763.0A Pending CN106011053A (en) | 2016-06-27 | 2016-06-27 | Novel dental pulp stem cell culturing medium |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106011053A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111411074A (en) * | 2019-01-05 | 2020-07-14 | 杭州可典生物科技有限公司 | Bone marrow cell culture medium |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103432007A (en) * | 2009-01-23 | 2013-12-11 | 首都医科大学 | New use of tooth-related stem cells |
| CN103849595A (en) * | 2012-12-05 | 2014-06-11 | 上海坤爱生物科技有限公司 | Large scale production technology for dental pulp stem cell |
| CN104630141A (en) * | 2015-02-03 | 2015-05-20 | 黑龙江天晴干细胞股份有限公司 | Method for preparing dental pulp mesenchymal stem cells and establishing dental pulp mesenchymal stem cell bank and cell resuscitation method |
| CN104694464A (en) * | 2015-02-09 | 2015-06-10 | 安徽新生命干细胞科技有限公司 | Clinical dental pulp stem cell and preparation method thereof |
| CN105087477A (en) * | 2015-05-21 | 2015-11-25 | 王松灵 | Application of mesenchymal stem cell modified by miR-21 antisense nucleotide |
-
2016
- 2016-06-27 CN CN201610475763.0A patent/CN106011053A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103432007A (en) * | 2009-01-23 | 2013-12-11 | 首都医科大学 | New use of tooth-related stem cells |
| CN103849595A (en) * | 2012-12-05 | 2014-06-11 | 上海坤爱生物科技有限公司 | Large scale production technology for dental pulp stem cell |
| CN104630141A (en) * | 2015-02-03 | 2015-05-20 | 黑龙江天晴干细胞股份有限公司 | Method for preparing dental pulp mesenchymal stem cells and establishing dental pulp mesenchymal stem cell bank and cell resuscitation method |
| CN104694464A (en) * | 2015-02-09 | 2015-06-10 | 安徽新生命干细胞科技有限公司 | Clinical dental pulp stem cell and preparation method thereof |
| CN105087477A (en) * | 2015-05-21 | 2015-11-25 | 王松灵 | Application of mesenchymal stem cell modified by miR-21 antisense nucleotide |
Non-Patent Citations (2)
| Title |
|---|
| AUBREY YOUNG: "Induction of Differentiation of Dental Pulp-Derived Mesenchymal Stem cells (DPSC)", 《HTTPS://DIGITALSCHOLARSHIP.UNLV.EDU》 * |
| KELCEY LOVELAND ET AL.: "Dental Pulp Stem Cell (DPSC) Pluripotency Enhanced by Transforming Growth Factor (TGF-β1) in Vitro may be Inhibited by Differentiation-Inducing Factors Laminin-5 and Dexamethasone", 《INTERNATIONAL JOURNAL OF BIOLOGICAL SCIENCES AND APPLICATIONS》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111411074A (en) * | 2019-01-05 | 2020-07-14 | 杭州可典生物科技有限公司 | Bone marrow cell culture medium |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Toupadakis et al. | Comparison of the osteogenic potential of equine mesenchymal stem cells from bone marrow, adipose tissue, umbilical cord blood, and umbilical cord tissue | |
| CN101210232B (en) | Mesenchyme stem cell preserving fluid | |
| CN103070161B (en) | Cryopreservation liquid and cryopreservation method for adipose tissue-derived mesenchymal stem cells ( ADSC) | |
| JP5775667B2 (en) | Methods of lithium stimulation of cord blood stem cell proliferation and growth factor production | |
| US20110002895A1 (en) | Composition for autotransplantation or allotransplantation using dental pulp stem cell, and use of the composition | |
| CN106414707A (en) | Method for producing culture medium composition | |
| CN102703385B (en) | Mesenchymal stem cell nutrient solution | |
| CN113564111B (en) | Method for culturing umbilical cord-derived mesenchymal stem cells in low-oxygen mode | |
| CN105079859A (en) | Dressing and preparation method thereof | |
| CN107502588B (en) | Method for separating and preparing dental pulp stem cells | |
| CN112481202A (en) | Method for serum-free separation culture of umbilical cord mesenchymal stem cells by using platelet lysate | |
| CN105505865A (en) | Separation method for umbilical cord mesenchymal stem cells | |
| CN106011053A (en) | Novel dental pulp stem cell culturing medium | |
| JP2011211956A (en) | Undifferentiation-maintaining agent for stem cell and growth-promoting agent | |
| KR102054645B1 (en) | Novel Mesenchymal stem cells derived from Tissue of Animal Tongue and Method for Preparing Thereof | |
| RU2645255C1 (en) | Method for obtaining of biosafe culture of mesenchimal stem cells from human chorionic villae | |
| JP5714267B2 (en) | Stem cell undifferentiation maintenance agent and proliferation promoter | |
| CN103805561A (en) | Culture medium for human mesenchymal stem cells | |
| CN104711308A (en) | Micromolecule substance for stimulating mesenchymal stem cells to secrete fibronectin | |
| JP5710145B2 (en) | Stem cell undifferentiation maintenance agent and proliferation promoter | |
| JP5710138B2 (en) | Stem cell undifferentiation maintenance agent and proliferation promoter | |
| JP2009232724A (en) | Stem cell cultured by specific method and method for producing the same | |
| CN116640727B (en) | Nutrient solution for improving cell viability and preparation method and application thereof | |
| CN108220232A (en) | A kind of cultural method of the culture medium based on fetal calf serum | |
| JP5710146B2 (en) | Stem cell undifferentiation maintenance agent and proliferation promoter |
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: 20161012 |
|
| RJ01 | Rejection of invention patent application after publication |