CN208218882U - A kind of combining ultrasonic targeted microbubble destroys the device of technology mediated gene transfection - Google Patents
A kind of combining ultrasonic targeted microbubble destroys the device of technology mediated gene transfection Download PDFInfo
- Publication number
- CN208218882U CN208218882U CN201820512372.6U CN201820512372U CN208218882U CN 208218882 U CN208218882 U CN 208218882U CN 201820512372 U CN201820512372 U CN 201820512372U CN 208218882 U CN208218882 U CN 208218882U
- Authority
- CN
- China
- Prior art keywords
- culture plate
- tissue culture
- destroys
- gene transfection
- mediated gene
- 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.)
- Expired - Fee Related
Links
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The utility model relates to the devices that a kind of combining ultrasonic targeted microbubble destroys technology mediated gene transfection, including lifting platform, iron stand, further include silica gel plug, ultrasonic probe, Ultrasound Transfection instrument and tissue culture plate；It is characterized in that, the lifting platform is placed on the iron stand pedestal；The ultrasonic probe is connected with Ultrasound Transfection instrument；The tissue culture plate is placed on lifting platform by the way of standing upside down；The tissue culture plate is sealed by silica gel plug；The silica gel plug is placed in immediately below tissue culture plate in isosceles triangle；The ultrasonic probe is placed in the surface of tissue culture plate；The ultrasonic probe is placed on iron clamp, and is fixed on iron stand.The utility model improves efficiency gene transfection, simplifies operating procedure, drains that air success rate is high, ensure that gene transfection safely, conveniently and efficiently.
Description
Technical field
The utility model relates to microculture instrument technical fields, and in particular to a kind of gene transfecting cell culture dress
It sets.
Background technique
Gene therapy is that normal gene or medicative inhereditary material are imported target cell by ad hoc fashion, to entangle
Orthogene defect plays therapeutic effect, to achieve the purpose that treat disease.In recent years, with the development of molecular biology,
Gene therapy shows unique advantage in terms of the treatment of major disease.Efficient gene transduction method is successfully to carry out base
Because of an important step for the treatment of, wherein the gene delivery systems of non-invasion, targeting clinically show particularly important.However base
The internal transport of cause is important problem in science during gene therapy, how by genetic safety, efficiently transport therapy section
Position is the significant challenge of gene therapy.Current main gene delivery system includes viral vectors and non-virus carrier.Virus carries
Body lack targeting so that gene be difficult to it is safe and efficient reach target site, cause curative effect undesirable, and since its is bright
Aobvious toxic side effect limits its clinical application.In non-virus carrier, lipid microbubble is by lipid outer membrane and inert gas core
Heart composition, can carry therapeutic gene, protect it from nuclease degradation in vivo, destroy in combining ultrasonic targeted microbubble
It, can be in specific time and space after (Ultrasound-targeted microbubble destruction, UTMD) technology
The load gene microvesicle in target tissue is smashed, therapeutic gene is discharged, and broadening vascular endothelial cell gap and of short duration increase are thin simultaneously
Gene transfection efficiency is improved to increase permeability of cell membrane in after birth hole.Therefore, UTMD is as a kind of ideal gene delivery side
Method has broad application prospects.
However in the research of UTMD rotaring redyeing gene, traditional positive placement tissue culture plate is inverted ultrasonic probe exposing cell
The problems such as obtained gene transfection efficiency of bottom plate is lower, and different document transfection efficiencies differ greatly.Based on this problem, it is necessary to existing
There is technology to improve.
Summary of the invention
It is traditional it is positive place tissue culture plate, be inverted the obtained gene transfection efficiency of ultrasonic probe exposing cell bottom plate compared with
It is low, the reason is that since the lipid microbubble of gas core easily swims in cell culture fluid surface, when forward direction places tissue culture plate,
The microvesicle and board bottom cell for carrying gene are there are certain distance, after ultrasound wave irradiation increases permeability of cell membrane, gene cannot and
When enter cell.
To solve the above-mentioned problems, the utility model provides a kind of combining ultrasonic targeted microbubble destruction technology mediation gene turn
The device of dye, tissue culture plate inversion can effectively improve gene transfection efficiency, provide safe and efficient gene delivery for gene therapy
Mode.
When tissue culture plate is inverted, microvesicle swims in cell peripheral, when ultrasound breaks up microvesicle, when open cells fenestra,
Gene is easier to enter intracellular.There is a necessary condition at this time, tissue culture plate must drain air after being inverted, otherwise air
It steeps oneself-meeting and floats up to cell peripheral, completely cut off microvesicle and cell, make the reduction of gene transfection efficiency instead.
The purpose of this utility model is achieved through the following technical solutions: a kind of combining ultrasonic targeted microbubble destruction technology
The device of mediated gene transfection, including lifting platform (5), iron stand (4) further include silica gel plug (7), ultrasonic probe (2), ultrasound turn
Contaminate instrument (1) and tissue culture plate (6)；It is characterized in that, the lifting platform (5) is placed on the iron stand (4) pedestal；Institute
Ultrasonic probe (2) is stated to be connected with Ultrasound Transfection instrument (1)；The tissue culture plate (6) is placed in lifting platform by the way of standing upside down
(5) on；The tissue culture plate (6) is sealed by silica gel plug (7)；The silica gel plug (7) is placed in tissue culture plate in isosceles triangle
(6) immediately below；The ultrasonic probe (2) is placed in the surface of tissue culture plate (6)；The ultrasonic probe (2) is placed in iron
It presss from both sides on (3), and is fixed on iron stand (4).
It is sealing structure when the silica gel plug (7) connect with tissue culture plate (6) in some technical solutions.
In some technical solutions, the ultrasonic probe (2) fits closely with tissue culture plate (6) bottom plate.
In some technical solutions, the tissue culture plate (6) is 12 orifice plates.
In some technical solutions, the iron clamp (3) is fixed on iron stand (4), and adjustable height and direction.
In some technical solutions, the tissue culture plate (6) uses non-toxic polyvinyl chloride board making.
In some technical solutions, the lifting platform (5) is using threaded rod control lifting.
In some technical solutions, the table top of the lifting platform (5) is equipped with anti-skidding silicagel pad.
In some technical solutions, the iron clamp (3) is circle, and one layer of rubber layer is wrapped up in outside.
The utility model uses silica gel plug encapsulated cell culture plate, fits closely silica gel plug with liquid level in cell hole, arranges
Air success rate is high to the greatest extent, easy to operate, guarantees the close contact of microvesicle and cell after tissue culture plate is inverted.
Iron stand of the utility model using lifting platform and adjustable height and direction, the essence of achievable cell hole and probe
Really fitting.
The utility model uses 12 orifice plates, and single hole is bonded completely with the ultrasonic probe shape, thin when can make ultrasound wave irradiation
Born of the same parents are uniform by shining, and ultrasonic wave is avoided to interfere other cell hole results.
Beneficial effect
It places tissue culture plate with conventional forward to carry out outside ultrasonic in combination microbubble body compared with cytogene transfection, this is practical new
Type improves efficiency gene transfection；Tissue culture plate, which is inverted, with other carries out cytogene transfection phase outside ultrasonic in combination microbubble body
Than the utility model in turn simplifies operating procedure, drains air success rate height, ensure that the safely, conveniently and high of gene transfection
Effect.
Detailed description of the invention
Fig. 1 is the utility model structure diagram.
In figure: 1, Ultrasound Transfection instrument；2, ultrasonic probe；3, iron clamp；4, iron stand；5, lifting platform；6, tissue culture plate；7,
Silica gel plug.
Specific embodiment
The utility model is described in further detail with reference to the accompanying drawing:
As shown in Figure 1, a kind of combining ultrasonic targeted microbubble destroys the device of technology mediated gene transfection, including lifting platform
(5), iron stand (4) further include silica gel plug (7), ultrasonic probe (2), Ultrasound Transfection instrument (1) and tissue culture plate (6)；The liter
Drop platform (5) is placed on the iron stand (4) pedestal；The ultrasonic probe (2) is connected with Ultrasound Transfection instrument (1)；It is described thin
Born of the same parents' culture plate (6) is placed on lifting platform (5) by the way of standing upside down；The tissue culture plate (6) is sealed by silica gel plug (7)
Mouthful；The silica gel plug (7) is placed in immediately below tissue culture plate (6) in isosceles triangle；The ultrasonic probe (2) is placed in cell culture
The surface of plate (6)；The ultrasonic probe (2) is placed on iron clamp (3), and is fixed on iron stand (4).
The operating procedure of the device is as follows:
(1) prepare on the day before ultrasound wave irradiation: under the conditions of 37 DEG C, 0.25% trypsin digestion cell 1-5min is obtained thin
Born of the same parents' suspension, by the plantation of its interval in 12 orifice plates, after cell incubator is placed for 24 hours, reaching 80%-90% then to cell density be can be used
In transfection.
(2) PBS is cleaned after cell for use, and self-control lipid microbubble, miRNA and serum free medium are incubated for 20min jointly, is added
Sample enters tissue culture plate.
(3) it is sealed using silica gel plug, air must be drained, then tissue culture plate is inverted in lifting by encapsulated cell hole
On platform.
(4) ultrasonic probe is fixed on iron stand by iron clamp.
(5) lifting platform and iron clamp are adjusted, makes ultrasonic probe and tissue culture plate bottom plate close contact, is emptied using couplant
Air, alignment need the cell hole irradiated.
(6) ultrasound wave irradiation continues culture 6-8 hours after irradiation, replace culture medium thereafter, and PBS is washed 3 times, removal
Remaining microvesicle, miRNA and dead cell.
Above embodiment is only preferred embodiments of the present invention, not only embodiment, for this skill
For the those of ordinary skill in art field, without departing from the principle of this utility model, can also make it is several improvement and
Retouching, these improvements and modifications are also considered as within the protection scope of the utility model.What is be not described in detail in this specification is interior
Appearance belongs to the prior art well known to professional and technical personnel in the field.
Claims (9)
1. a kind of combining ultrasonic targeted microbubble destroys the device of technology mediated gene transfection, including lifting platform (5), iron stand (4),
It further include silica gel plug (7), ultrasonic probe (2), Ultrasound Transfection instrument (1) and tissue culture plate (6)；It is characterized in that, the lifting
Platform (5) is placed on the iron stand (4) pedestal；The ultrasonic probe (2) is connected with Ultrasound Transfection instrument (1)；The cell
Culture plate (6) is placed on lifting platform (5) by the way of standing upside down；The tissue culture plate (6) is sealed by silica gel plug (7)；
The silica gel plug (7) is placed in immediately below tissue culture plate (6) in isosceles triangle；The ultrasonic probe (2) is placed in tissue culture plate
(6) surface；The ultrasonic probe (2) is placed on iron clamp (3), and is fixed on iron stand (4).
2. a kind of combining ultrasonic targeted microbubble according to claim 1 destroys the device of technology mediated gene transfection, special
Sign is, is sealing structure when the silica gel plug (7) connect with tissue culture plate (6).
3. a kind of combining ultrasonic targeted microbubble according to claim 1 destroys the device of technology mediated gene transfection, special
Sign is that the ultrasonic probe (2) fits closely with tissue culture plate (6) bottom plate.
4. a kind of combining ultrasonic targeted microbubble according to claim 1 destroys the device of technology mediated gene transfection, special
Sign is that the tissue culture plate (6) is 12 orifice plates.
5. a kind of combining ultrasonic targeted microbubble according to claim 1 destroys the device of technology mediated gene transfection, special
Sign is that the iron clamp (3) is fixed on iron stand (4), and adjustable height and direction.
6. a kind of combining ultrasonic targeted microbubble according to claim 1 or 4 destroys the device of technology mediated gene transfection,
It is characterized in that, the tissue culture plate (6) uses non-toxic polyvinyl chloride board making.
7. a kind of combining ultrasonic targeted microbubble according to claim 1 destroys the device of technology mediated gene transfection, special
Sign is that the lifting platform (5) is using threaded rod control lifting.
8. a kind of combining ultrasonic targeted microbubble according to claim 1 destroys the device of technology mediated gene transfection, special
Sign is that the table top of the lifting platform (5) is equipped with anti-skidding silicagel pad.
9. a kind of combining ultrasonic targeted microbubble according to claim 1 destroys the device of technology mediated gene transfection, special
Sign is that the iron clamp (3) is circle, and one layer of rubber layer is wrapped up in outside.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201820512372.6U CN208218882U (en) | 2018-04-10 | 2018-04-10 | A kind of combining ultrasonic targeted microbubble destroys the device of technology mediated gene transfection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201820512372.6U CN208218882U (en) | 2018-04-10 | 2018-04-10 | A kind of combining ultrasonic targeted microbubble destroys the device of technology mediated gene transfection |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN208218882U true CN208218882U (en) | 2018-12-11 |
Family
ID=64514803
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201820512372.6U Expired - Fee Related CN208218882U (en) | 2018-04-10 | 2018-04-10 | A kind of combining ultrasonic targeted microbubble destroys the device of technology mediated gene transfection |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN208218882U (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109652456A (en) * | 2019-01-22 | 2019-04-19 | 北京大学深圳医院 | Receptor transfection microvesicle, ligand transfection microvesicle and two sections of conduction cell transfecting methods processed for cell transfecting |
| CN109706183A (en) * | 2019-01-15 | 2019-05-03 | 深圳市圣祥高科技有限公司 | A kind of Ultrasound Transfection device |
| WO2022021734A1 (en) * | 2020-07-27 | 2022-02-03 | 中国科学院深圳先进技术研究院 | Ultrasonic sound field conformal gene transfection device |
-
2018
- 2018-04-10 CN CN201820512372.6U patent/CN208218882U/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109706183A (en) * | 2019-01-15 | 2019-05-03 | 深圳市圣祥高科技有限公司 | A kind of Ultrasound Transfection device |
| CN109652456A (en) * | 2019-01-22 | 2019-04-19 | 北京大学深圳医院 | Receptor transfection microvesicle, ligand transfection microvesicle and two sections of conduction cell transfecting methods processed for cell transfecting |
| WO2022021734A1 (en) * | 2020-07-27 | 2022-02-03 | 中国科学院深圳先进技术研究院 | Ultrasonic sound field conformal gene transfection device |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN208218882U (en) | A kind of combining ultrasonic targeted microbubble destroys the device of technology mediated gene transfection | |
| Du et al. | Advanced physical techniques for gene delivery based on membrane perforation | |
| Kinoshita et al. | A novel method for the intracellular delivery of siRNA using microbubble-enhanced focused ultrasound | |
| Zarnitsyn et al. | Physical parameters influencing optimization of ultrasound-mediated DNA transfection | |
| Liang et al. | Optimisation of ultrasound-mediated gene transfer (sonoporation) in skeletal muscle cells | |
| CN202610237U (en) | Operating platform beneficial for ultrasound mediating gene transfection | |
| Fan et al. | Inhibition of prostate cancer growth using doxorubicin assisted by ultrasound-targeted nanobubble destruction | |
| Manome et al. | Insonation facilitates plasmid DNA transfection into the central nervous system and microbubbles enhance the effect | |
| Jinturkar et al. | Gene delivery using physical methods | |
| Sun et al. | Development of therapeutic microbubbles for enhancing ultrasound-mediated gene delivery | |
| CN105779386B (en) | A kind of application of mescenchymal stem cell in preparation treatment M5 type leukemia medicament | |
| CN106031795A (en) | Ultrasound-microbubble transferring device and method thereof | |
| Zhang et al. | Enhancement effect of ultrasound-induced microbubble cavitation on branched polyethylenimine-mediated VEGF165 transfection with varied N/P ratio | |
| US20020009706A1 (en) | Method for loading cells with an agent | |
| Sancho-Albero et al. | Use of exosomes as vectors to carry advanced therapies | |
| Deng et al. | Improving the efficacy of therapeutic angiogenesis by UTMD-mediated Ang-1 gene delivery to the infarcted myocardium | |
| Li et al. | Sonoporation: underlying mechanisms and applications in cellular regulation | |
| Wang et al. | Enhanced homing ability and retention of bone marrow stromal cells to diabetic nephropathy by microbubble-mediated diagnostic ultrasound irradiation | |
| Bekeredjian et al. | Impact of microbubbles on shock wave-mediated DNA uptake in cells in vitro | |
| US20070196283A1 (en) | Composition for transfection of DNA into the liver | |
| CN101475962B (en) | Method for mediated gene transfection | |
| CN106635793A (en) | Culture dish for ultrasonic experiment and use method | |
| CN111254162B (en) | Preparation of cationic lipid microvesicles and mediated gene delivery method thereof | |
| WO2022021734A1 (en) | Ultrasonic sound field conformal gene transfection device | |
| CN113999814A (en) | Method for inhibiting human breast cancer cells by external electric field and interferometer |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20181211 Termination date: 20210410 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |