CN109337864A - A kind of preparation method of dextrorotation hydrogel material - Google Patents
A kind of preparation method of dextrorotation hydrogel material Download PDFInfo
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- CN109337864A CN109337864A CN201811358395.7A CN201811358395A CN109337864A CN 109337864 A CN109337864 A CN 109337864A CN 201811358395 A CN201811358395 A CN 201811358395A CN 109337864 A CN109337864 A CN 109337864A
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- C12N5/06—Animal cells or tissues; Human cells or tissues
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- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
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- C12N2506/13—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells from connective tissue cells, from mesenchymal cells
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- C12N2506/1353—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells from connective tissue cells, from mesenchymal cells from mesenchymal stem cells from bone marrow mesenchymal stem cells (BM-MSC)
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Abstract
The present invention relates to a kind of preparation methods of dextrorotation hydrogel material, which solve existing host materials to be difficult to the technical issues of accuracy controlling stem cell breaks up at rouge, its key step are as follows: (1) dextrorotation gelator is dissolved in dimethyl sulphoxide solution, the dextrorotation gelator solution that mass-volume concentration is 12mg/ul~33mg/ul is obtained, 24 orifice plate bottoms are placed in；(2) dextrorotation gelator solution obtained is mixed into the culture medium suspension of mesenchymal stem cell, is mixed in 24 orifice plates, 30~60min is stood under the conditions of 30~40 DEG C, forms dextrorotation hydrogel；(3) dextrorotation hydrogel obtained is put into the mescenchymal stem cell culture medium culture of no Osteoinductive Factor, interval time replaces the mescenchymal stem cell culture medium on the dextrorotation hydrogel.It the composite can be widely applied to aquagel fibre and regulate and control three-dimensional mescenchymal stem cell into rouge differentiation field.
Description
Technical field
The present invention relates to biological chemical field, specifically a kind of preparation method of dextrorotation hydrogel material.
Background technique
The adjustable cell fate of growth local microenvironment and cell behavior of stem cell, and guide growth course.In embryo
During fetal hair is educated, extracellular matrix microenvironment participates in regulation embryo molding and organ occurs.The physical environment tune of multipotential stem cell
Save their self-renewing and differentiation.Mechanical and physics clue is also critically important in adult tissue, and wherein adult stem cell needs
Physical interaction with extracellular matrix is to maintain its effect.Therefore, how stem cell is adjusted by extracellular matrix to break up
And function, become nowadays regenerative medicine research hotspot.From bionical and tissue repair demand angle design and the high biology of building
Active scaffold material realizes the selective regulation of histocyte function, promotes bone tissue reparation to rebuild important as medical domain
Developing direction.Therefore, stem cell response timbering material microenvironment feature, which causes function differentiation, is the core of problem in science, to set
Meter and the timbering material with " biological response regulation " function.
Summary of the invention
The present invention is exactly to be difficult to the technical issues of accuracy controlling stem cell breaks up at rouge to solve existing host material, is mentioned
A kind of preparation method of dextrorotation hydrogel material that the interaction regulation stem cell using cell and material breaks up at rouge is supplied.
For this purpose, the present invention provides a kind of preparation methods of dextrorotation hydrogel material, comprising the following steps:
(1) dextrorotation gelator is dissolved in dimethyl sulphoxide solution, acquisition mass-volume concentration is 12mg/ul~33mg/
The dextrorotation gelator solution of ul, is placed in 24 orifice plate bottoms；
(2) dextrorotation gelator solution obtained in step (1) is mixed into the culture medium suspension of mesenchymal stem cell,
It is mixed in 24 orifice plates, 30~60min is stood under the conditions of 30~40 DEG C, form dextrorotation hydrogel；
(3) dextrorotation hydrogel obtained in step (2) is put into the mescenchymal stem cell culture medium of no Osteoinductive Factor
Culture, interval time replace the mescenchymal stem cell culture medium on dextrorotation hydrogel.
Preferably, dextrorotation gelator is the symmetrical phenylalanine derivative class hydrogelator of C2.
Preferably, the number containing mesenchymal stem cell is 100,000 in culture medium suspension.
Preferably, interval time is 2 days.
Preferably, the mesenchymal stem cell in dextrorotation hydrogel can be grown 7 days or more in right-handed chirality environment.
Helpfulness of the invention:
The present invention design one kind can with dimensional culture stem cell hydrogel material, and can by change material molecule it is chiral this
One fundamental characteristics achievees the purpose that regulate and control stem cell destiny, solves existing host material and be difficult to accuracy controlling stem cell into rouge
Break up this technological difficulties, to meet the needs of clinical precisely medical treatment.
In the present invention, regulating and controlling mesenchymal stem cell into the material that rouge breaks up is dextrorotation gelator, specifically, C2 pairs
The phenylalanine derivative class hydrogelator of title can be self-assembly of fibrous web-like chain in cell suspension, be the growth of cell
The porous environment of three-dimensional fiber, the response using cell to three-dimensional space chirality are provided, dextrorotation fibre space structure can promote ring
Domestic mesenchymal stem cell breaks up at rouge, and dextrorotation hydrogel mixing stem cell implantation subcutaneous area has apparent fat raw
At.
Detailed description of the invention
Fig. 1 is the dextrorotation fibre space that the symmetrical phenylalanine derivative class hydrogelator of C2 in the present invention is formed
Scanning electron microscope；
Fig. 2 (a) is after 1 dextrorotation hydrogel culture of the embodiment of the present invention 7 days, and immunofluorescence observation is detected at rouge；Fig. 2 (b)
After 2 dextrorotation hydrogel culture of the embodiment of the present invention 7 days, immunofluorescence observation is detected at rouge.
Specific embodiment
According to following embodiments, the present invention may be better understood.However, as it will be easily appreciated by one skilled in the art that real
It applies content described in example and is merely to illustrate the present invention, without this hair described in claims should will not be limited
It is bright.
Embodiment 1
It is dissolved in dimethyl sulphoxide solution using dextrorotation gelator, it is solidifying to obtain the dextrorotation that mass-volume concentration is 12mg/ul
Glue factor solutions, are placed in 24 orifice plate bottoms, and the culture medium suspension containing 100,000 mesenchymal stem cells of 500ul is quickly infused
It injects in 24 above-mentioned orifice bores, dextrorotation hydrogel can be formed by standing 60 minutes at 40 DEG C.
The mescenchymal stem cell culture medium without Osteoinductive Factor is added in dextrorotation hydrogel, and (group is divided into mescenchymal stem cell
Fetal calf serum+100IU/mL the Pen .- Strep of basal medium+10% is bought in Sai Ye Biotechnology Co., Ltd,
Similarly hereinafter), during culture, at interval of the mescenchymal stem cell culture medium on 2 days replacement dextrorotation hydrogels.
After dextrorotation hydrogel mixing mescenchymal stem cell culture 3 days, it is implanted into subcutaneous rat region.
After dextrorotation hydrogel mixing mescenchymal stem cell culture 7 days, the sight of mesenchymal stem cell immunofluorescence is carried out
It examines into Adipose Differentiation to detect, such as Fig. 2 (a).
Embodiment 2
It is dissolved in dimethyl sulphoxide solution using dextrorotation gelator, it is solidifying to obtain the dextrorotation that mass-volume concentration is 23mg/ul
Glue factor solutions, are placed in 24 orifice plate bottoms, and the culture medium suspension containing 100,000 mesenchymal stem cells of 500ul is quickly infused
It injects in 24 above-mentioned orifice bores, dextrorotation hydrogel can be formed by standing 45 minutes at 35 DEG C.
The mescenchymal stem cell culture medium without Osteoinductive Factor is added in dextrorotation hydrogel, during culture, at interval of 2 days
Replace the mescenchymal stem cell culture medium on dextrorotation hydrogel.
After dextrorotation hydrogel mixing mescenchymal stem cell culture 3 days, it is implanted into subcutaneous rat region.
After dextrorotation hydrogel mixing mescenchymal stem cell culture 7 days, the sight of mesenchymal stem cell immunofluorescence is carried out
It examines into Adipose Differentiation to detect, such as Fig. 2 (b).
Embodiment 3
It is dissolved in dimethyl sulphoxide solution using dextrorotation gelator, it is solidifying to obtain the dextrorotation that mass-volume concentration is 33mg/ul
Glue factor solutions, are placed in 24 orifice plate bottoms, and the culture medium suspension containing 100,000 mesenchymal stem cells of 500ul is quickly infused
It injects in 24 above-mentioned orifice bores, dextrorotation hydrogel can be formed by standing 30 minutes at 30 DEG C.
The mescenchymal stem cell culture medium culture without Osteoinductive Factor is added in dextrorotation hydrogel, wherein without osteogenic induction
Stem cell media is supplemented between the factor with money, during culture, at interval of the mescenchymal stem cell culture on 2 days replacement dextrorotation hydrogels
Base.
After dextrorotation hydrogel mixing mescenchymal stem cell culture 3 days, it is implanted into subcutaneous rat region.
After dextrorotation hydrogel mixing mescenchymal stem cell culture 7 days, mesenchymal stem cell is carried out into Adipose Differentiation
Detection.
Conclusion: by Examples 1 to 3 it is found that the symmetrical phenylalanine derivative class hydrogelator of C2 can be outstanding in cell
It is self-assembly of fibrous web-like chain in liquid, provides three-dimensional fiber porous environment for the growth of cell, using cell to three-dimensional space
Chiral response, dextrorotation fibre space structure can promote the mesenchymal stem cell in environment to break up at rouge, dextrorotation water-setting
Glue mixing stem cell implantation subcutaneous area has apparent fat to generate.
Claims (5)
1. a kind of preparation method of dextrorotation hydrogel material, characterized in that comprise the steps of:
(1) dextrorotation gelator is dissolved in dimethyl sulphoxide solution, obtaining mass-volume concentration is 12mg/ul~33mg/ul's
Dextrorotation gelator solution is placed in 24 orifice plate bottoms；
(2) dextrorotation gelator solution obtained in the step (1) is mixed into the culture medium suspension of mesenchymal stem cell,
It is mixed in 24 orifice plates, 30~60min is stood under the conditions of 30~40 DEG C, form dextrorotation hydrogel；
(3) dextrorotation hydrogel obtained in the step (2) is put into the mescenchymal stem cell culture medium of no Osteoinductive Factor
Culture, interval time replace the mescenchymal stem cell culture medium on the dextrorotation hydrogel.
2. the preparation method of dextrorotation hydrogel material according to claim 1, which is characterized in that in step (1), the right side
Rotation gelator is the symmetrical phenylalanine derivative class hydrogelator of C2.
3. the preparation method of dextrorotation hydrogel material according to claim 1, which is characterized in that described in step (2)
The number containing mesenchymal stem cell is 100,000 in culture medium suspension.
4. the preparation method of dextrorotation hydrogel material according to claim 1, which is characterized in that in step (3), between described
It is 2 days every the time.
5. the preparation method of dextrorotation hydrogel material according to claim 1, which is characterized in that in step (3), the right side
Mesenchymal stem cell in rotation hydrogel can be grown 7 days or more in right-handed chirality environment.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115389365A (en) * | 2022-10-31 | 2022-11-25 | 北京大学口腔医学院 | Method for evaluating the properties of supramolecular hydrogel carriers |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103694131A (en) * | 2013-12-06 | 2014-04-02 | 上海交通大学 | Chiral supermolecule hydrogel and preparation method and application thereof |
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- 2018-11-15 CN CN201811358395.7A patent/CN109337864A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103694131A (en) * | 2013-12-06 | 2014-04-02 | 上海交通大学 | Chiral supermolecule hydrogel and preparation method and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| TAMMAY DAS等: "phenylalanine and derivatives as versatile low-molecular-weight gelators:design,structure and tailored function", 《BIOMATERIALS SCIENCE》 * |
| 江圣杰等: "三维手性微环境调控干细胞命运研究", 《第十三次全国老年口腔医学学术年会论文汇编》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115389365A (en) * | 2022-10-31 | 2022-11-25 | 北京大学口腔医学院 | Method for evaluating the properties of supramolecular hydrogel carriers |
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