CN105462835A - Double-layer assembly type cytomechanics loading carrier device - Google Patents
Double-layer assembly type cytomechanics loading carrier device Download PDFInfo
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- CN105462835A CN105462835A CN201410445006.XA CN201410445006A CN105462835A CN 105462835 A CN105462835 A CN 105462835A CN 201410445006 A CN201410445006 A CN 201410445006A CN 105462835 A CN105462835 A CN 105462835A
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Abstract
The invention relates to a carrier device for conducting tensile stress loading on cells, and the device is composed of an upper layer clamping clamp, an intermediate layer clamping clamp, a lower layer clamping clamp, multiple screws, an upper layer silicone rubber membrane and a lower layer silicone rubber membrane. The area of a cell culture basement membrane is increased through the clamping clamps and the screws, meanwhile, double layer design is adopted, and the effective culture area of each stressed cell is greatly increased. By means of the double-layer assembly type cytomechanics loading carrier device, effective mechanical loading can be conducted on the four layers of cells (the front side and the reverse side of the basement membrane) at the same time, and the requirement of the high throughput test for the number of cells is satisfied. The clamps, the screws and the basement membrane are all made of transparent materials, and the biocompatibility is excellent. Real-time microscopical cell morphology observation can be achieved. The whole device is simple and fast in assembling process and cell inoculation process, after being assembled, the device is fitted to all types of existing mechanical loading equipment with axial tension as the design principle as a whole, and loading and unloading are both convenient and efficient. The whole carrier device can be sterilized once again and used repeatedly, the experimental process is simplified, and experiment efficiency is greatly improved.
Description
Technical field
The present invention relates to a kind of carrier arrangement cell being implemented to Tensile stress loading, be related specifically to and need to apply mechanical stimulation to a large amount of cell, to solve in relevant cell biomechanics Research, carrier Effective Mechanical Properties load-carrying area is less, cell harvesting comparatively small amt, can not reach the problem that high throughput testing requires.
Background technology
Along with cytobiology, the fast development of biomechanics and biomaterial, by biomechanical environment in analogue body, make observation mechanical stimulation on cell proliferation, the research of differentiation and cell physiological function becomes possibility, the impact of mechanics microenvironment on cell also more and more comes into one's own, and there is multiple mechanics loading regime, as four-point bending loads, substrate tension loads, and liquid or gas static pressure load, centrifugal force and hydrodynamic shear etc., also devise multiple mechanics load apparatus simultaneously.Existing Tensile stress loading regime, mainly rely on the elastic substrates film of tensile cell growth attachment, make basilar membrane generation deformation, thus power is conducted to cell, and pass through the size and the frequency that control basilar membrane distortion, thus realize implementing effective mechanical stimulation to cell.Patent CN1425905 has invented a kind of four-point bending Mechanical loading instrument, and by the power of stepper-motor, two eccentric point of contact on sheathed screw, realize the upwards deformation to culture plate above, thus realize the Mechanical loading to cell.But the different point of contact spacing of its culture plate is different, and the unbalance stress of whole culture plate, all can not ensure that cell is subject to identical mechanical stimulation.Patent CN101092595A is invented and is a kind ofly changed the elastic substrates film cultivated and have cell by mechanical top board moving up and down, to realize the mechanical stimulation to cell.The same with four-point bending principle, also there is the unity and homogeneity that can not ensure Mechanical loading in this design.Patent CN1932511A devises a kind of single shaft sinusoidal tensile mechanical loading unit, by realizing the mechanical stimulation to cell to the axial tension of elastica, although the loading needs compared with large sstrain amount can be met, but once can only implement to load to a cell culture unit, have impact on its scope of application and Efficiency.
Also all there is shortcoming and defect in above-mentioned existing patent: the culture plate one, loaded or the basilar membrane area of stretching less, had a strong impact on the results quantity of target cell, cannot high throughput testing have been realized, as real-time quantitative PCR and gene microarray analysis etc.Two, cell loading unit can not be independent, observes under being unfavorable for the real time microscopic of handling and cell.Three, basilar membrane or cell carrier device etc., the process being installed to Mechanical loading equipment is also comparatively complicated, also there is the sterilizing of basilar membrane simultaneously, cell inoculation is all inconvenient.What above deficiency was serious limits the using and promoting in cell biomechanics research field of elastic substrates film.
Summary of the invention
The present invention be directed to the above weak point of existing Tensile stress deceleration loading device, and a kind of general cyto-mechanics proposed loads carrier arrangement.Apparatus of the present invention, by polycarbonate clamping fixture and screw, are widened the area of cell cultures basilar membrane, are adopted double layer design simultaneously, add the culture area of stressed cell greatly.Two layers of substrate film can be loaded simultaneously, once can cultivate face to four confluent monolayer cells, implement Mechanical loading (pros and cons of basilar membrane), solve high throughput testing to the high order of magnitude requirement of cell.Wherein clamping fixture, screw and basilar membrane are transparent material, no cytotoxicity, and biocompatibility is excellent.Under transparent substrates film can realize real-time microscope, cell routine dyes and morphological observation.The assembling of whole device and inoculating cell process simple and direct, be fitted to as a whole with axial tension after assembling be principle of design existing various Mechanical loading equipment on, simultaneously can high pressure steam, radioactive rays or ethylene oxide sterilizing, convenient and efficient.
The present invention is achieved by the following technical solutions:
1, be made up of upper strata clamping fixture, middle level clamping fixture and lower floor's clamping fixture, some screws and upper layer of silicon rubber diaphragm, lower floor's silicone rubber membrane.
2, upper strata clamping fixture, middle level clamping fixture and lower floor's clamping fixture and screw, be polycarbonate material, jointly completes the fixing of whole device.
3, upper strata clamping fixture, middle level clamping fixture and lower floor's clamping fixture each a pair, is long strip type.By adjusting the length of clamping fixture, the size adjusting the effective load-carrying area of silicone rubber membrane can be realized.
4, the centre of upper strata clamping fixture and lower floor's clamping fixture, is designed with foursquare clamp area; Middle level clamping fixture is without clamp area.
5, the clamp area of upper strata clamping fixture and lower floor's clamping fixture, carries out the process of intersection rag.To increase clamping friction power, make it more firm by what clamp on mechanics equipment.
6, the two ends of upper strata clamping fixture, middle level clamping fixture and lower floor's clamping fixture, are provided with the clamping fixture screw supporting with screw.
7, upper layer of silicon rubber diaphragm and lower floor's silicone rubber membrane are rectangular flake.Silicone rubber membrane is transparent material, and biocompatibility is good.Directly cell dyeing and morphological observation can be carried out under microscope.
8, four angles of upper layer of silicon rubber diaphragm and lower floor's silicone rubber membrane, in the position that respective screws is passed, are provided with the silicone rubber membrane screw with screw mates.
Accompanying drawing explanation
Accompanying drawing: Fig. 1: structural representation of the present invention.
Fig. 2: upper strata of the present invention clamping fixture, middle level clamping fixture and lower floor's clamping fixture schematic diagram.
Fig. 3: upper layer of silicon rubber diaphragm of the present invention and lower floor's silicone rubber membrane schematic diagram.
Fig. 4: side of the present invention is seen.
Fig. 5: anterior aspect of the present invention.
Fig. 6: sight above of the present invention.
In figure: 1. lower floor's clamping fixture; 2. middle level clamping fixture; 3. upper strata clamping fixture; 4. screw; 5. upper layer of silicon rubber diaphragm; 6. lower floor's silicone rubber membrane; 7. clamp area; 8. silicone rubber membrane screw; 9. clamping fixture screw.
Embodiment
The present invention is suitable for the cyto-mechanics loading equipemtn of existing various axial tension principle design, is made up of upper strata clamping fixture, middle level clamping fixture and lower floor's clamping fixture, some screws and upper layer of silicon rubber diaphragm, lower floor's silicone rubber membrane.Before using, first by the thoroughly cleaning respectively of each several part assembly, for subsequent use after dry.Get a pair lower floor's clamping fixture, clamp area keeps flat downwards, and lower floor's silicone rubber membrane is lain on lower floor's clamping fixture, and aims at corresponding screw.Again middle level clamping fixture is overlayed on lower floor's silicone rubber membrane, and aim at corresponding screw.Then continue to stack upper layer of silicon rubber diaphragm, and aim at corresponding screw.Finally upper strata clamping fixture with position upwards, clamp area, be superimposed upon on upper layer of silicon rubber diaphragm, and aim at corresponding screw.After having superposed, fixed with screw is top-down.While tightening screw, adjust the position of each clamping fixture and silicone rubber membrane, to ensure that each clamping fixture and silicone rubber membrane are all in smooth state of unfolding, thus whole device and the uniform force of silicone rubber membrane when ensureing Mechanical loading.Sterilizing (high pressure steam, ray or ethylene oxide sterilizing) is carried out to the integral carriers device after assembling, for subsequent use.
Prepare to need the some milliliters of cell suspension of inoculation (inoculum density and the order of magnitude experimentally itself determine) for subsequent use.Integral carriers after sterilizing is put in advance the sterile petri dish of suitable diameter and the degree of depth, inoculate appropriate cell suspension (about 2ml) respectively on the silicone rubber membrane of the upper and lower, the liquid tension on film surface is relied on to keep suspension to be uniformly distributed in silicone rubber membrane surface, culture dish and carrier are moved in constant incubator together, cultivates 2-4 hour.After cell major part is adherent, take out culture dish and integral carriers device, integral carriers device is overturn, the upper and lower silicone rubber membrane is put upside down mutually.In kind the surface of the two panels silicone rubber membrane of cell suspension inoculation after upset, then put back to incubator cultivate 2-4 hour, until cell complete adherent after, in culture dish, add substratum, be advisable not have whole culture carrier device.Cell cultures routine changes liquid, and inverted microscope observation of cell proliferative conditions, take out after reaching the cell density of requirement of experiment, and integral carriers is assembled in corresponding cyto-mechanics loading equipemtn, the Mechanical loading of axial tension can be implemented.
Claims (8)
1. double-deck sectional cyto-mechanics loads a carrier arrangement, is made up of upper strata clamping fixture, middle level clamping fixture and lower floor's clamping fixture, some screws and upper layer of silicon rubber diaphragm and lower floor's silicone rubber membrane.
2. double-deck sectional cyto-mechanics according to claim 1 loads carrier arrangement, it is characterized in that: use the upper strata clamping fixture of polycarbonate material, middle level clamping fixture and lower floor's clamping fixture and screw, the assembling having carried out whole device is fixed.
3. double-deck sectional cyto-mechanics according to claim 1 loads carrier arrangement, and it is characterized in that: upper strata clamping fixture, middle level clamping fixture and lower floor's clamping fixture, be long strip type.
4. double-deck sectional cyto-mechanics according to claim 3 loads carrier arrangement, it is characterized in that: have a foursquare clamp area in the middle of upper strata clamping fixture and lower floor's clamping fixture.
5. double-deck sectional cyto-mechanics according to claim 4 loads carrier arrangement, it is characterized in that: the clamp area of upper strata clamping fixture and lower floor's clamping fixture, carries out the process of intersection rag.
6. double-deck sectional cyto-mechanics according to claim 1 loads carrier arrangement, and it is characterized in that: upper strata clamping fixture, the two ends of middle level clamping fixture and lower floor's clamping fixture are provided with the clamping fixture screw with screw mates.
7. double-deck sectional cyto-mechanics according to claim 1 loads carrier arrangement, it is characterized in that: upper layer of silicon rubber diaphragm and lower floor's silicone rubber membrane are rectangular flake.
8. double-deck sectional cyto-mechanics according to claim 1 loads carrier arrangement, it is characterized in that: four angles of upper layer of silicon rubber diaphragm and lower floor's silicone rubber membrane, in the position that respective screws is passed, is provided with the silicone rubber membrane screw with screw mates.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410445006.XA CN105462835A (en) | 2014-09-03 | 2014-09-03 | Double-layer assembly type cytomechanics loading carrier device |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410445006.XA CN105462835A (en) | 2014-09-03 | 2014-09-03 | Double-layer assembly type cytomechanics loading carrier device |
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| CN105462835A true CN105462835A (en) | 2016-04-06 |
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| CN201410445006.XA Pending CN105462835A (en) | 2014-09-03 | 2014-09-03 | Double-layer assembly type cytomechanics loading carrier device |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105199955A (en) * | 2015-10-22 | 2015-12-30 | 重庆大学 | Three-dimensional cell culture device capable of performing mechanical-chemical coupling loading |
| CN112608845A (en) * | 2021-01-11 | 2021-04-06 | 兰州大学第一医院 | Bionic stimulation device for mechanical pressure of stem cells |
-
2014
- 2014-09-03 CN CN201410445006.XA patent/CN105462835A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105199955A (en) * | 2015-10-22 | 2015-12-30 | 重庆大学 | Three-dimensional cell culture device capable of performing mechanical-chemical coupling loading |
| CN112608845A (en) * | 2021-01-11 | 2021-04-06 | 兰州大学第一医院 | Bionic stimulation device for mechanical pressure of stem cells |
| CN112608845B (en) * | 2021-01-11 | 2023-07-25 | 兰州大学第一医院 | Stem cell mechanical pressure bionic stimulation device |
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Application publication date: 20160406 |
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| WD01 | Invention patent application deemed withdrawn after publication |