CN107082893A - A kind of gelatine microsphere and preparation method thereof - Google Patents
A kind of gelatine microsphere and preparation method thereof Download PDFInfo
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- CN107082893A CN107082893A CN201710318416.1A CN201710318416A CN107082893A CN 107082893 A CN107082893 A CN 107082893A CN 201710318416 A CN201710318416 A CN 201710318416A CN 107082893 A CN107082893 A CN 107082893A
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- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J3/00—Processes of treating or compounding macromolecular substances
- C08J3/24—Crosslinking, e.g. vulcanising, of macromolecules
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J3/00—Processes of treating or compounding macromolecular substances
- C08J3/02—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques
- C08J3/03—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques in aqueous media
- C08J3/05—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques in aqueous media from solid polymers
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- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
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- C08K5/15—Heterocyclic compounds having oxygen in the ring
- C08K5/151—Heterocyclic compounds having oxygen in the ring having one oxygen atom in the ring
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- C08L89/00—Compositions of proteins; Compositions of derivatives thereof
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
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Abstract
The present invention relates to field of cell culture, more particularly to a kind of gelatine microsphere and preparation method thereof.The invention provides a kind of preparation method of gelatine microsphere, step is：Gelatin is soluble in water, different emulsifier for mixing are separately added into, emulsion is obtained；Different dehydrating agents will be added in emulsion, are cleaned, after drying, sieving, gelatine microsphere crude product are obtained；The ethanol solution containing Geniposide crosslinking and curing agent is added in the gelatine microsphere crude product, gelatine microsphere is prepared.Test result indicates that, toxic action of the gelatine microsphere to cell made from the preparation method provided by the present invention is smaller, and the toxicity to cell can be reduced during cell amplification cultivation, improves cytoactive, and degradation effect is preferable.
Description
Technical field
The present invention relates to field of cell culture, more particularly to a kind of gelatine microsphere and preparation method thereof.
Background technology
Gelatin is the good high polymer material of natural origin, biocompatibility, is collagen through gentle and irreversible drop
The product that solution is obtained.Contain keratin, elasticin, melanocyte and chondroitin in gelatin, sticking and raw for cell can be promoted
Long, this is that gelatin can expand a key factor of microcarrier as cell.When temperature is down to 40 DEG C or so, gelatin can occur by
The transformation to screw type conformation is freely crimped, therefore gelatin can not be used directly in physiological conditions, want to expand as cell
Carrier, then must carry out appropriate crosslinking.
At present, there is certain toxic side effect to cell in conventional aldehyde crosslinking agent (such as glutaraldehyde), be unfavorable for cell
Amplification.
The content of the invention
In view of this, object of the present invention is to provide a kind of gelatine microsphere and preparation method thereof, what the present invention was provided
Gelatine microsphere can reduce the toxicity to cell during cell amplification cultivation, improve cytoactive, and degradation effect is preferable.
The present invention provides a kind of preparation method of gelatine microsphere, comprises the following steps：
A) it is gelatin is soluble in water, dissolve by heating, the oil phase added dissolved with emulsifying agent A is stirred, and adds emulsifying agent B
It is stirred, rapid cooling is again stirring for, obtains emulsion.
B) dehydrating agent A is added in the emulsion to be dehydrated, add dehydration de-oiling agent B and dehydration de-oiling agent C is handed over
For cleaning gelatine microsphere, finally with washes of absolute alcohol, after drying, sieving, gelatine microsphere crude product is obtained.
C) ethanol solution containing Geniposide crosslinking and curing agent is added in the gelatine microsphere crude product, a period of time is placed
Afterwards, Aspirate supernatant, sequentially passes through cleaning, drying, obtains gelatine microsphere.
It is preferred that, step a) the emulsifying agent A are atoleine, castor oil, olive oil, peanut oil, soybean oil, cocoa bean
Fat, cocounut oil vinegar, palm fibre put any one or a few in oily vinegar, ethyl acetate, beeswax and silicone oil.
It is preferred that, step a) the emulsifying agent B are atoleine, castor oil, olive oil, peanut oil, soybean oil, cocoa
Beans fat, cocounut oil vinegar, palm fibre put any one or a few in oily vinegar, ethyl acetate, beeswax and silicone oil.
It is preferred that, it is characterised in that step b) the dehydrating agent A are absolute ethyl alcohol, isopropanol, acetone, glycerine, n-butanol
And any one in the tert-butyl alcohol.
It is preferred that, step b) the dehydration de-oiling agent B are absolute ethyl alcohol, isopropanol, acetone, glycerine, dioxanes, positive fourth
Any one in alcohol, the tert-butyl alcohol.
It is preferred that, step b) the dehydration de-oiling agent C are absolute ethyl alcohol, isopropanol, acetone, glycerine, dioxanes, positive fourth
Any one in alcohol, the tert-butyl alcohol.
It is preferred that, mass volume ratio of step c) the Geniposide crosslinking and curing agents in ethanol solution be 0.25%~
1%.
It is preferred that, the particle diameter of step c) the gelatine microsphere crude products is 75~300 μm.
The present invention also provides a kind of gelatine microsphere, is prepared by preparation method described in above-mentioned any one.
Compared with prior art, the present invention provides a kind of gelatine microsphere and preparation method thereof.The gelatin that the present invention is provided is micro-
Geniposide crosslinking and curing agent is added in the preparation method of ball.The Geniposide (genipin) of natural origin be a class glycosaminoglycan/
The good crosslinking agent of protein, its toxic action to cell is significantly less than glutaraldehyde, can subtract during cell amplification cultivation
Few toxicity to cell, improves cytoactive, and degradation effect is preferable.
Brief description of the drawings
In order to illustrate more clearly about the embodiment of the present invention or technical scheme of the prior art, below will be to embodiment or existing
There is the accompanying drawing used required in technology description to be briefly described, it should be apparent that, drawings in the following description are only this
Some embodiments of invention, for those of ordinary skill in the art, without having to pay creative labor, may be used also
To obtain other accompanying drawings according to these accompanying drawings.
Fig. 1 shows the schematic diagram of cell attachment gelatine microsphere 1；
Fig. 2 shows the schematic diagram of cell attachment gelatine microsphere 2；
Fig. 3 shows the schematic diagram of cell attachment gelatine microsphere 3；
Fig. 4 shows the schematic diagram of cell attachment gelatine microsphere 4；
Fig. 5 shows the degraded situation schematic diagram of gelatine microsphere 1；
Fig. 6 shows the degraded situation schematic diagram of gelatine microsphere 2；
Fig. 7 shows the degraded situation schematic diagram of gelatine microsphere 3；
Fig. 8 shows the degraded situation schematic diagram of gelatine microsphere 4.
Embodiment
Below in conjunction with the accompanying drawing in the embodiment of the present invention, the technical scheme in the embodiment of the present invention is carried out clear, complete
Site preparation is described, it is clear that described embodiment is only a part of embodiment of the invention, rather than whole embodiments.It is based on
Embodiment in the present invention, it is every other that those of ordinary skill in the art are obtained under the premise of creative work is not made
Embodiment, belongs to the scope of protection of the invention.
The invention provides a kind of preparation method of gelatine microsphere, comprise the following steps：
First, it is gelatin is soluble in water.The gelatin consumption is preferably 10%-20%w/v.It is dissolved in progress heating after water molten
Solution.The heating is preferably 70 DEG C -100 DEG C.The oil phase added dissolved with emulsifying agent A is stirred.The emulsifying agent A is preferably liquid
Body paraffin, castor oil, olive oil, peanut oil, soybean oil, cocoa butter, cocounut oil vinegar, palm fibre put oily vinegar, ethyl acetate, beeswax and
It is any in any one or a few in silicone oil, more preferably atoleine, ethyl acetate, olive oil, peanut oil, soybean oil
It is one or more of.Gelatin volume and emulsifying agent A volume ratio are preferably 1~5:1, it is stirred with certain speed, the speed
Degree is preferably 100r/min~900r/min, and emulsifying agent B is added after stirring and is stirred.The emulsifying agent B is preferably liquid
Paraffin, castor oil, olive oil, peanut oil, soybean oil, cocoa butter, cocounut oil vinegar, palm fibre put oily vinegar, ethyl acetate, beeswax and silicon
It is any one in any one or a few in oil, more preferably atoleine, ethyl acetate, olive oil, peanut oil, soybean oil
Plant or several.The addition of the emulsifying agent B is that the volume ratio of gelatine microsphere and emulsifying agent B is 1:1~5.Then by emulsion
Ice-water bath is rapidly cooled to less than 5 DEG C, continues to stir a period of time with certain speed, obtains emulsion.The speed of the stirring
Degree is preferably 100r/min~450r/min.
Then, the dehydrating agent A that -20 DEG C of ice baths are added in the emulsion is dehydrated.The addition of the dehydrating agent
For 2~5 times of emulsion volume.After dehydration terminates, adding dehydration de-oiling agent B and dehydration de-oiling agent C, alternately cleaning gelatin is micro-
Ball.The dehydration de-oiling agent B is preferably one kind of absolute ethyl alcohol, isopropanol, acetone, glycerine, dioxanes, n-butanol, the tert-butyl alcohol,
More preferably isopropanol, acetone, dioxanes.The dehydration de-oiling agent C is preferably absolute ethyl alcohol, isopropanol, acetone, glycerine, two
Oxane, n-butanol, one kind of the tert-butyl alcohol, more preferably isopropanol, acetone, dioxanes.Finally washed again with absolute ethyl alcohol
Wash, place and dried under certain temperature, obtain gelatin microspheres granules.The drying temperature is preferably 60 DEG C~80 DEG C.Will after drying
The gelatine microsphere is sieved.The screen mesh size of the sieving is respectively 75 μm, 160 μm, 200 μm, 300 μm.After sieving
Obtain the gelatine microsphere crude product of different-grain diameter.
Finally, the gelatine microsphere crude product of certain particle diameter is chosen, the ethanol solution containing Geniposide crosslinking and curing agent is added.Institute
The particle diameter for stating gelatine microsphere is preferably 75 μm~300 μm.The addition of the ethanol solution containing Geniposide crosslinking and curing agent
For 2~5 times of gelatine microsphere crude product volume, the mass volume ratio of the Geniposide crosslinking and curing agent in the solution is 0.25%~
1%.20~30h is placed at 4 DEG C, 36~40h is then placed at 37 DEG C, supernatant is abandoned in suction.Add washes of absolute alcohol 2-3
Secondary, drying obtains gelatine microsphere.The addition of the absolute ethyl alcohol is preferably 2~5 times of gelatine microsphere crude product.
The present invention also provides a kind of gelatine microsphere, is prepared by the preparation method of above-mentioned gelatine microsphere.
Compared with prior art, the present invention provides a kind of gelatine microsphere and preparation method thereof.The gelatin that the present invention is provided is micro-
Geniposide crosslinking and curing agent is added in the preparation method of ball.The Geniposide (genipin) of natural origin be a class glycosaminoglycan/
Olefinic carbon atoms on the good crosslinking agent of protein, the gelatin mechanism of crosslinking with uniqueness, Geniposide are by amino on gelatin
Nucleophillic attack, open loop forms heterocyclic amine compound, and its toxic action to cell is significantly less than glutaraldehyde.In cell amplification cultivation
During can reduce the toxicity to cell, improve cytoactive, and degradation effect is preferable.
Current Geniposide has been obtained for widely studied and application as Biological cross-linker in biomedical sector, successfully
Applied to various aspects such as artificial creature's dressing, organizational project cornea, oesophagus, tracheae, articular cartilages.It is de- that Geniposide is crosslinked
Cell bovine pericardium has more preferable biocompatibility than glutaraldehyde cross-linking effect, and cytotoxicity is low, and structure stability is high, more
Plus it is suitable as valvular tissue engineering scaffold material.Geniposide contains multiple active function groups such as hydroxyl, carboxyl, therefore can be used for
The crosslinking of a variety of natural biologic materials, its cross-linking products formed have the characteristics of stabilization, anti-inflammatory, promotion organization regenerate.
To enable goal of the invention, feature, the advantage of the present invention more obvious and understandable, below in conjunction with the present invention
Accompanying drawing in embodiment, the technical scheme in the embodiment of the present invention is clearly and completely described, it is clear that disclosed below
Embodiment be only a part of embodiment of the invention, and not all embodiment.Based on the embodiment in the present invention, this area
All other embodiment that those of ordinary skill is obtained under the premise of creative work is not made, belongs to protection of the present invention
Scope.
Embodiment 1 prepares gelatine microsphere
(1) 5g gelatin is weighed, is dissolved in ultra-pure water 50mL, is heated and be completely dissolved at 70 DEG C.Gelatin solution stirring is added
Dissolved with the oil phase of emulsion liquid paraffin.The volume ratio of gelatin volume and emulsion liquid paraffin is 5:1, with 900r/min's
Speed is stirred, and forms emulsion.Emulsifying agent olive oil is added, second layer oil phase is formed.Gelatin volume and emulsifying agent olive oil
Volume ratio be 1:2,1min is stirred with 450rpm speed immediately.
(2) its speed is made to be cooled to less than 5 degrees Celsius emulsion ice-water bath, 450rpm speed continues to stir 30min, then
Add the dehydration de-oiling agent absolute ethyl alcohol of -20 DEG C of ice baths of 2 times of emulsion volumes, stirring dehydration.Again with dehydration de-oiling reagent
Isopropanol, acetone alternately clean gelatine microsphere, finally wash gelatine microsphere from absolute ethyl alcohol, place 80 DEG C of drying.Gelatin is micro-
Ball particle is respectively with 75 μm, and 300 μm of screen clothes sieve successively.
(3) select the gelatine microsphere of 75 μm~300 μm of size, add 5 times of volumes containing the golden Buddhist nun of 0.25% crosslinking agent
Flat ethanol solution.4 DEG C of 30h are placed, are then placed 36 hours at 37 DEG C.Supernatant is abandoned in suction, by gelatine microsphere with 5 times of volumes
Washes of absolute alcohol 2-3 times, 100 degrees Celsius of drying gelatine microspheres 1.
Embodiment 2 prepares gelatine microsphere
(1) 5g gelatin is weighed, is dissolved in ultra-pure water 50mL, is completely dissolved in 70 DEG C of heating.Gelatin solution stirring is added
Dissolved with the oil phase of emulsifying agent ethyl acetate.The volume ratio of gelatin volume and emulsifying agent ethyl acetate is 2:1, with 900r/min's
Speed is stirred, and forms emulsion.Emulsifying agent soybean oil is added, second layer oil phase is formed.Gelatin volume and emulsifying agent soybean oil
Volume ratio be 1:3.1min is stirred with 450rpm speed immediately.
(2) emulsion ice-water bath is then quickly cooled to less than 5 degrees Celsius, 450rpm speed continues to stir 30min.
Add the dehydration de-oiling agent isopropanol of -20 DEG C of ice baths of 2 times of emulsion volumes, stirring dehydration.Again with dehydration de-oiling reagent
Isopropanol, acetone alternately clean gelatine microsphere, finally wash gelatine microsphere from absolute ethyl alcohol, place 80 DEG C of drying.Gelatin is micro-
Ball particle is respectively with 75 μm, and 300 μm of screen clothes sieve successively
(3) gelatine microsphere of 75 μm -300 μm of size is selected, add 5 times of volumes contains 0.6% crosslinking agent Jin Niping
Ethanol solution.20h is placed at 4 DEG C, is then placed 30 hours at 37 DEG C.Supernatant is abandoned in suction, by gelatine microsphere with 5 times of bodies
Long-pending washes of absolute alcohol 2-3 times, 100 degrees Celsius of drying gelatine microspheres 2.
Embodiment 3 prepares gelatine microsphere
(1) 10g gelatin is weighed, is dissolved in ultra-pure water 50mL, is completely dissolved in 70 DEG C of heating.Gelatin solution stirring adds
Enter in the oil phase dissolved with emulsifying agent ethyl acetate.The volume ratio of gelatin volume and emulsifying agent ethyl acetate is 1:1, with 900r/min
Speed stirring, formed emulsion.Emulsifying agent soybean oil is added, second layer oil phase is formed.Gelatin volume and emulsifying agent peanut
The volume ratio of oil is 1:3. 1min is stirred with 200rpm speed immediately.
(2) emulsion ice-water bath is then quickly cooled to less than 5 degrees Celsius, 450rpm speed continues to stir 30min.
Add the dehydration de-oiling agent isopropanol of -20 DEG C of ice baths of 2 times of emulsion volumes, stirring dehydration.Again with dehydration de-oiling reagent
Dioxanes, isopropanol alternately clean gelatine microsphere, finally wash gelatine microsphere from absolute ethyl alcohol, place 80 DEG C of drying.Gelatin
Microsphere particle is respectively with 75 μm, and 300 μm of screen clothes sieve successively.
(3) gelatine microsphere of 75 μm -300 μm of size is selected, add 5 times of volumes contains 1% crosslinking agent Jin Niping's
90% ethanol solution.4 DEG C of 20h are placed, are then placed 30 hours at 37 DEG C.Supernatant is abandoned in suction, by nothing of the gelatine microsphere with 5 times of volumes
Water-ethanol is cleaned 2-3 times, 100 degrees Celsius of drying gelatine microspheres 3.
The comparative example of embodiment 4
(1) 10g gelatin is weighed, is dissolved in ultra-pure water 50mL, is completely dissolved in 70 DEG C of heating.Gelatin solution stirring is added
Dissolved with the oil phase of emulsifying agent ethyl acetate.The volume ratio of gelatin volume and emulsifying agent ethyl acetate is 1:1, with 900r/min's
Speed is stirred, and forms emulsion.Emulsifying agent soybean oil is added, second layer oil phase is formed.Gelatin volume and emulsifying agent peanut oil
Volume ratio be 1:3.
(2) 1min is stirred with 200rpm speed immediately.Then by emulsion ice-water bath be quickly cooled to 5 degrees Celsius with
Under, 450rpm speed continues to stir 30min.It is 10% to add glutaraldehyde to its content, and crosslinking curing 3 hours adds 2 times
The dehydration de-oiling agent isopropanol of -20 DEG C of ice baths of the emulsion volume, stirring dehydration.Again with dehydration de-oiling reagent dioxanes, different
Propyl alcohol alternately cleans gelatine microsphere
(3) gelatine microsphere finally is washed from absolute ethyl alcohol, places 80 DEG C of drying.Gelatin microspheres granules respectively with 75 μm,
300 μm of screen clothes sieve successively, obtain control group gelatine microsphere 4.
Embodiment 5 uses gelatine microsphere expanding stem cells
Gelatine microsphere prepared by embodiment 1~4 is after autoclave sterilization, with DMEM/F12 basal medium water
Change, be placed on 37 DEG C of activation and stay overnight.The mixing of UC-MSCs P2 umbilical cord mesenchymal stem cells is inoculated with, is trained with DMEM/F12+10%FBS
Foster base is seeded in 24 orifice plates, and static to place 37 DEG C, 5%CO2 incubators co-culture 48 hours, obtain cell suspension.
Embodiment 6DAPI dyeing detection cell attachment gelatine microsphere situations
Supernatant is abandoned in the gelatine microsphere suspension of Example 1~4, suction, is added DAPI and is dyed 15 seconds, after PBS 2 times, glimmering
Taken pictures on light microscope and observe the situation of cell attachment gelatine microsphere, Fig. 1~Fig. 4 shows that cell distinguishes adherent gelatine microsphere 1~4.
As a result show (table 1, cell have adherent gelatine microsphere for+, acellular adherent gelatine microsphere for -), use gelatin
Microballoon 1, gelatine microsphere 2, gelatine microsphere 3 cell attachment effect preferably, the adherent effect of comparative example gelatine microsphere 4 is general.
The cell attachment result of table 1
| Gelatine microsphere 1 | Gelatine microsphere 2 | Gelatine microsphere 3 | Gelatine microsphere 4 | |
| Cell attachment | +++ | +++ | +++ | + |
The detection of cell viability after the degraded of the gelatine microsphere of embodiment 7
The gelatine microsphere suspension of Example 1~4, draws supernatant, after PBS gelatine microsphere 2-3 times, adds 0.25%
Pancreas enzyme -EDTA, is resuspended gelatine microsphere, is placed on 37 degrees Celsius of incubation 15-30min, is shaken once per 10min, until gelatin is micro-
Ball is degradable, adds FBS and stops digestion, and with 1000rpm centrifuge cell suspension 5min, suction abandons supernatant, uses DMEM/F12+10%
Precipitation is resuspended in FBS complete mediums, and cell suspension is carried out into trypan blue, carries out cell viability detection.
The cell viability of table 2 is detected
| Gelatine microsphere 1 | Gelatine microsphere 2 | Gelatine microsphere 3 | Gelatine microsphere 4 | |
| Cell viability | 90.2% | 85% | 89.5% | 80% |
As shown in Table 2, the cell viability after gelatine microsphere 1,2,3 is cultivated can reach more than 85%, gelatine microsphere 1,2,3
Between cell viability indifference (P>0.5), cell can keep good vigor, right because crosslinking agent glutaraldehyde has toxic side effect
Cell viability after ratio gelatine microsphere 4 is cultivated only reaches 80%, the cell of gelatine microsphere 1,2,3 and comparative example gelatine microsphere 4
Vigor has differences (P<0.05), cell viability is poor.
The degraded of the gelatine microsphere of embodiment 8
Gelatine microsphere prepared by Example 1~4, adds 0.25% pancreas enzyme -EDTA, after mixing, and gelatine microsphere is resuspended, puts
Put and be incubated 30min at 37 degrees Celsius, concussion once, adds FBS and stops digestion, in Microscopic observation gelatin microspheres granules per 10min
Degraded situation simultaneously counts granule number.
The gelatine microsphere of table 3 degraded remainder particulate
| Gelatine microsphere 1 | Gelatine microsphere 2 | Gelatine microsphere 3 | Gelatine microsphere 4 | |
| Granule number | 0 | 0 | 2 | >200 |
Degradation effect such as Fig. 5~Fig. 8 shows that the degradation effect of gelatine microsphere 1,2,3 is preferable, substantially without residual particles, gelatin
Microballoon 4 still has abundant residues.As known from Table 3, indifference (P between gelatine microsphere 1,2,3>0.05), and gelatine microsphere 1,2,3 with
Between comparative example gelatine microsphere 4, there is notable difference (P<0.01).
Described above, the above embodiments are merely illustrative of the technical solutions of the present invention, rather than its limitations；Although with reference to before
Embodiment is stated the present invention is described in detail, it will be understood by those within the art that：It still can be to preceding
State the technical scheme described in each embodiment to modify, or equivalent substitution is carried out to which part technical characteristic；And these
Modification is replaced, and the essence of appropriate technical solution is departed from the spirit and scope of various embodiments of the present invention technical scheme.
Claims (9)
1. a kind of preparation method of gelatine microsphere, comprises the following steps：
A) it is gelatin is soluble in water, dissolve by heating, the oil phase added dissolved with emulsifying agent A is stirred, add emulsifying agent B progress
Stirring, rapid cooling, is again stirring for, obtains emulsion.
B) dehydrating agent A is added in the emulsion to be dehydrated, add dehydration de-oiling agent B and dehydration de-oiling agent C alternatings are clear
Gelatine microsphere is washed, finally with washes of absolute alcohol, after drying, sieving, gelatine microsphere crude product is obtained.
C) ethanol solution containing Geniposide crosslinking and curing agent is added in the gelatine microsphere crude product, is placed after a period of time,
Supernatant is abandoned in suction, is sequentially passed through cleaning, drying, is obtained gelatine microsphere.
2. preparation method according to claim 1, it is characterised in that step a) the emulsifying agent A are atoleine, castor-oil plant
Oil, olive oil, peanut oil, soybean oil, cocoa butter, cocounut oil vinegar, palm fibre put appointing in oily vinegar, ethyl acetate, beeswax and silicone oil
Meaning is one or more of.
3. preparation method according to claim 1, it is characterised in that step a) the emulsifying agent B are atoleine, castor-oil plant
Oil, olive oil, peanut oil, soybean oil, cocoa butter, cocounut oil vinegar, palm fibre put appointing in oily vinegar, ethyl acetate, beeswax and silicone oil
Meaning is one or more of.
4. preparation method according to claim 1, it is characterised in that step b) the dehydrating agent A are absolute ethyl alcohol, isopropyl
Any one in alcohol, acetone, glycerine, n-butanol and the tert-butyl alcohol.
5. preparation method according to claim 1, it is characterised in that step b) the dehydration de-oiling agent B are absolute ethyl alcohol, different
Any one in propyl alcohol, acetone, glycerine, dioxanes, n-butanol, the tert-butyl alcohol.
6. preparation method according to claim 1, it is characterised in that step b) the dehydration de-oiling agent C are absolute ethyl alcohol, different
Any one in propyl alcohol, acetone, glycerine, dioxanes, n-butanol, the tert-butyl alcohol.
7. preparation method according to claim 1, it is characterised in that step c) the Geniposide crosslinking and curing agents are molten in ethanol
Mass volume ratio in liquid is 0.25%~1%.
8. preparation method according to claim 1, it is characterised in that the particle diameter of step c) the gelatine microsphere crude products is 75~
300μm。
9. a kind of gelatine microsphere, it is characterised in that prepared by preparation method described in claim 1 to 8 any one.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108753700A (en) * | 2018-04-02 | 2018-11-06 | 广州赛莱拉干细胞科技股份有限公司 | A kind of method of umbilical cord mesenchymal stem cells induction chondroblast |
| CN109627786A (en) * | 2018-12-21 | 2019-04-16 | 西北大学 | The preparation method of gelatine microsphere and its application in colored drawing class historical relic's protection |
| CN109793717A (en) * | 2017-11-17 | 2019-05-24 | 中国科学院大连化学物理研究所 | A kind of preparation method and application of gelatine microsphere |
| CN112250892A (en) * | 2020-10-22 | 2021-01-22 | 苏州新丝原生物科技有限公司 | Gelatin microsphere and preparation method and application thereof |
| WO2021211569A1 (en) * | 2020-04-13 | 2021-10-21 | Massachusetts Institute Of Technology | Dissolvable gelatin-based microcarriers generated through droplet microfluidics for expansion and culture of mesenchymal stromal cell |
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| CN108753700A (en) * | 2018-04-02 | 2018-11-06 | 广州赛莱拉干细胞科技股份有限公司 | A kind of method of umbilical cord mesenchymal stem cells induction chondroblast |
| CN109627786A (en) * | 2018-12-21 | 2019-04-16 | 西北大学 | The preparation method of gelatine microsphere and its application in colored drawing class historical relic's protection |
| WO2021211569A1 (en) * | 2020-04-13 | 2021-10-21 | Massachusetts Institute Of Technology | Dissolvable gelatin-based microcarriers generated through droplet microfluidics for expansion and culture of mesenchymal stromal cell |
| CN112250892A (en) * | 2020-10-22 | 2021-01-22 | 苏州新丝原生物科技有限公司 | Gelatin microsphere and preparation method and application thereof |
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