CN104498350B - A kind of homogenizer for separating subcellular components - Google Patents
A kind of homogenizer for separating subcellular components Download PDFInfo
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- CN104498350B CN104498350B CN201410772165.0A CN201410772165A CN104498350B CN 104498350 B CN104498350 B CN 104498350B CN 201410772165 A CN201410772165 A CN 201410772165A CN 104498350 B CN104498350 B CN 104498350B
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M1/00—Apparatus for enzymology or microbiology
- C12M1/24—Apparatus for enzymology or microbiology tube or bottle type
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M1/00—Apparatus for enzymology or microbiology
- C12M1/33—Disintegrators
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M1/00—Apparatus for enzymology or microbiology
- C12M1/34—Measuring or testing with condition measuring or sensing means, e.g. colony counters
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M1/00—Apparatus for enzymology or microbiology
- C12M1/36—Apparatus for enzymology or microbiology including condition or time responsive control, e.g. automatically controlled fermentors
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M1/00—Apparatus for enzymology or microbiology
- C12M1/36—Apparatus for enzymology or microbiology including condition or time responsive control, e.g. automatically controlled fermentors
- C12M1/38—Temperature-responsive control
Abstract
The invention discloses a kind of homogenizer for separating subcellular components, cell or tissue block can be prepared into the homogenate comprising various subcellular components, described homogenizer includes homogenate tube, homogenate tube includes tube wall and is positioned at the tube core of tube wall, described tube wall top seal, pipe lid is stamped in bottom, described tube core includes stator sleeve pipe and is arranged in the rotor of sleeve pipe, described pipe covers and is provided with rotor connection, rotor is connected with rotor connection, described homogenizer also includes the motor being positioned at outside homogenate tube, and motor provides rotary power by rotor connection for rotor.Described homogenate tube can be designed to tissue homogenate pipe or cell homogenates pipe as required, conveniently switch use, the present invention uses the automatic system with special tube core, microprocessor defines special application program by rule of operation optimization, make sample preparation procedure standardization, sequencing, reduce the impact of anthropic factor, obtain subcellular structure homogenate, separate subcellular structure for density gradient centrifugation with differential centrifugation.
Description
Technical field
The present invention relates to the separation of subcellular structure, particularly to a kind of homogenizer for separating subcellular components.
Background technology
Cell is made up of various subcellular structures, including nucleus, mitochondrion, lysosome, autophagosome, endoplasmic reticulum, Golgi body etc., observe these subcellular structures or carry out biochemical function analysis, it is necessary to separating the various subcellular components of purification, centrifugation technique is the basic means realizing this target.Before centrifugal, it is necessary to first prepare cell homogenates or tissue homogenate.Homogenate is under cryogenic, being placed in homogenizer by cell or tissue and add isotonic homogenate medium (i.e. 0.25mol/L sucrose 0.003mol/L calcium chloride solution) grinding, making cell or tissue mechanically be ground becomes the mixture comprising various subcellular components and inclusion.
The mode of homogenate has multiple: manual homogenization, machine homogenate, ultrasonic homogenate, multigelation etc., and what animal vegetable tissue or cell can carry out softness homogenizes process, obtains tissue homogenate or cell homogenates that experiment needs.
Manual homogenization is to pour into tissue pieces or cell together with homogenate medium or normal saline to carry out homogenate in glass homogenate tube, left hand is held homogenate tube and is inserted lower end in the vessel filling mixture of ice and water, stamp stem is inserted perpendicularly in sleeve pipe by the right hand, rotate upwardly and downwardly tens of times of grinding (6～8 minutes), fully grind, make tissue homogenate.Use glass homogenizer manual homogenization both to require great effort, time-consuming, uniform not again, also add the probability of pollution.Machine homogenate, is utilize lapping principle, and employing machine is power, drives homogenate bar to move in homogenizer, and what animal vegetable tissue carried out softness homogenizes process, obtains cell homogenates or tissue homogenate that experiment needs.Although manpower has been liberated in machine homogenate, it is still open-sky technique, it is easy to pollute.Ultrasonic Pulverization, is make cell breakage with ultrasonic cell disruption instrument, it is easy to damaging cells device, and can affect the activity of some enzyme.The cell cultivated or separate can with above method homogenate, repeatedly freeze-thaw about 3 times (can also namely allow cell add appropriate hypotonic medium or distilled water is lowerd and frozen in temperature refrigerator, dissolve, freeze again, re-dissolved, about 3 times repeatedly), but have the part enzyme activity can be impacted, and organelle release is not exclusively.Summary of the invention
The present invention seeks to: a kind of tissue for separating subcellular structure or cell homogeniser, preparation tissue easy, quick, efficient or cell homogenates are provided.
The technical scheme is that
A kind of homogenizer for separating subcellular components, cell or tissue block is separated into subcellular structure, described homogenizer includes homogenate tube, described homogenate tube includes tube wall and is positioned at the tube core of tube wall, described tube wall top seal, pipe lid is stamped in bottom, described tube core includes stator sleeve pipe and is arranged in the rotor of sleeve pipe, described pipe covers and is provided with rotor connection, rotor is connected with rotor connection, described separator also includes the motor being positioned at outside homogenate tube, and described motor provides rotary power by rotor connection for rotor.
A kind of preferred version is: be divided into Archimedian screw area under control and homogenate milling zone in the tube core of described Potter-Elvehjem Tissue Grinders from top to bottom, described Archimedian screw area under control is built with screw rod, homogenate milling zone is built with grinding rod, described screw rod and grinding rod connect to form rotor, connecting rotor connection bottom grinding rod, the position of the stator sleeve inside pipe wall correspondence grinding rod of described homogenate milling zone is provided with grinding layer.Described stator sleeve pipe base portion is provided with some equally distributed holes.It is further preferred that the diameter of described hole is 2.0-3.0mm.
Another kind of preferred version is: be divided into Archimedian screw area under control, homogenate milling zone and negative pressure rotary-cut district in the tube core of described cell homogenates pipe from top to bottom, described Archimedian screw area under control is built with screw rod, homogenate milling zone is built with grinding rod, negative pressure rotary-cut district is built with rotary-cut cutter head, described screw rod, grinding rod and rotary-cut cutter head are sequentially connected with composition rotor, connect rotor connection bottom rotary-cut cutter head.The position of the stator sleeve inside pipe wall correspondence grinding rod of described homogenate milling zone is provided with grinding layer.The part of the stator sleeve pipe correspondence rotary-cut cutter head in described negative pressure rotary-cut district is provided with some equally distributed holes.It is further preferred that the diameter of described hole is 0.8-1.2mm.
For above two scheme:
Preferably, described rotor is sealed by the joint of rotor connection with pipe lid.
Preferably, it is additionally provided with protection thermal insulation cover outside described homogenate tube.
It is further preferred that described protection thermal insulation cover is built with temperature probe and refrigeration compressor, described temperature probe and refrigeration compressor are connected with a microprocessor respectively, and described refrigeration compressor is also associated with circulating fan；Described microprocessor is also connected to motor and a touch screen.
The invention have the advantage that
1. the homogenizer for separating subcellular structure provided by the present invention, with the use of tissue homogenate pipe or cell homogenates pipe, built-in special tube core, composition automatic system, microprocessor defines special application program (time, speed, direction) by rule of operation optimization, it is possible to processes sample easily, makes sample preparation procedure standardization, sequencing, reducing the impact of anthropic factor, change operator's dependency is non-operator dependency.
2. the homogenizer of the present invention can select different homogenate tubes as required, when sample is piece of tissue, can adopt with the trizonal tissue homogenate pipe in Archimedian screw area under control, homogenate milling zone and negative pressure rotary-cut district, can directly select the cell homogenates pipe with Archimedian screw area under control, two regions of homogenate milling zone when sample is single cell suspension, switching uses flexibly.During using-system homogenate tube, required tissue samples only need to simply prepare into the piece of tissue of several millimeters size before separation, and the peeling knife in tissue homogenate pipe can be peeled into 1.0mm automatically3The tissue fragments of size, and then prepare into subcellular structure homogenate by homogenate milling zone, can be used for density gradient centrifugation and separate subcellular structure with differential centrifugation.
3. the present invention is by setting specific program, controls speed, time and direction, present invention may also apply to prepare histone homogenate.
4. the present invention has protection thermal insulation cover, and control realization protection insulation case temperature, at 0-4 DEG C, is protected organelle and protein function, provided sealed aseptic working environment simultaneously, ensure user's safety in utilization.
5. the present invention operates gentleness, and easy, quick, efficient.
Accompanying drawing explanation
Below in conjunction with drawings and Examples, the invention will be further described:
Fig. 1 is the structural principle of the homogenizer for separating subcellular structure of the present invention；
Fig. 2 is the structural representation of tissue homogenate pipe of the present invention；
Fig. 3 is the exploded view of tissue homogenate pipe shown in Fig. 2；
Fig. 4 is the structural representation of cell homogenates pipe of the present invention；
Fig. 5 is the exploded view of cell homogenates pipe shown in Fig. 4；
Fig. 6 is the schematic diagram of temperature probe of the present invention.
Detailed description of the invention
As it is shown in figure 1, the disclosed homogenizer for piece of tissue or cell homogenates being separated into subcellular structure, including homogenate tube, homogenate tube is arranged below motor, motor speed is adjustable, can reverse, and can operate according to set program (time, speed, direction).Protection thermal insulation cover it is additionally provided with outside described homogenate tube; described protection thermal insulation cover is built with temperature probe, refrigeration compressor and circulating fan; described temperature probe, refrigeration compressor and circulating fan are connected with a microprocessor respectively; realize in protection thermal insulation cover temperature automatically controlled 0-4 DEG C, protection organelle and protein function.Microprocessor is also connected with motor, controls rotary speed and the direction of motor, described microprocessor is also connected with a touch screen, is used for inputting situation in operational factor and display running.
Embodiment 1
As shown in Figures 2 and 3, provide a kind of tissue homogenate pipe, when sample is piece of tissue, this tissue homogenate pipe can be adopted, described tissue homogenate pipe, including tube wall 1 and the tube core 2 being positioned at tube wall, described tube wall top seal, pipe lid 3 is stamped in bottom, described tube core 2 includes stator sleeve pipe 4 and is arranged in the rotor 5 of sleeve pipe 4, described pipe lid 3 is provided with rotor connection, rotor is connected with rotor connection, described rotor is sealed by the joint of rotor connection with pipe lid, motor below homogenate tube provides rotary power by rotor connection for rotor, stator sleeve pipe 4 and tube wall are fixed on motor by bayonet socket.
Described tube core 2 is divided into Archimedian screw area under control, homogenate milling zone and negative pressure rotary-cut district from top to bottom, and the stator casing part correspondence negative pressure rotary-cut district of tube core is provided with the hole 6 that some equally distributed diameters are 0.8-1.2mm.Described Archimedian screw area under control is built with screw rod 51, homogenate milling zone is built with grinding rod 52, negative pressure rotary-cut district is built with rotary-cut cutter head 53, and described screw rod 51, grinding rod 52 and rotary-cut cutter head 53 are sequentially connected with composition rotor 5, connects rotor connection bottom rotary-cut cutter head 53.
Wherein: Archimedian screw area under control: the screw rod structure of rotor collectively constitutes Archimedian screw pipe with stator sleeve pipe, sleeve pipe cylinder internal coiling round is utilized to rise and promote liquid, its operation principle is when internal coiling rotates, screw rod rotates around the axis of itself on the one hand, it rolls along sleeve inner surface again on the other hand, then forms the sealing chamber of liquid.Screw rod each rotation, the water in annular seal space pushes ahead a pitch, and with the continuous rotation of work screw rod, water presses to another annular seal space from an annular seal space, is finally extruded from cannula tip by liquid, meanwhile, forms negative pressure at sleeve bottom, causes suction.
Negative pressure rotary-cut district: the suction that negative pressure is formed makes piece of tissue be close to casing wall, and sinks into hole, and the tissue cutting being absorbed in from cavity is got off by the rotary-cut cutter head of high speed rotating, forms 1.0mm3The tissue fragments of size.
Homogenate milling zone: tissue fragments, under the extracting effect of negative pressure, rises with liquid stream, through homogenate milling zone, the polytetrafluoroethylene grinding layer grinding being ground the polytetrafluoroethylene homogenate work surface on rod and tube core inwall is broken for subcellular structure homogenate.
Liquid in homogenate tube carries tissue fragments and constantly enters from the aperture of stator sleeve bottom, respectively through negative pressure rotary-cut district, homogenate milling zone and Archimedian screw area under control, flowing out from cannula tip, so go round and begin again, tissue grinder is broken for subcellular structure homogenate the most at last.In tissue homogenate program, design has Umklapp process, when rotor reverses, and the liquid reversal of direction of flow in tube core, the hole in tissue pieces blocking negative pressure rotary-cut district can be avoided.
The sample quality scope that the present invention separates: 20-4000mg；Rotor speed: 50-1000rpm, can rotate by both forward and reverse directions；Time: 1-600 second；Temperature: temperature controllable is in 0-4 DEG C；Application program: require to provide the rule of operation optimized and homogenate program according to different piece of tissue features and homogenate；Sample disposal: be aseptic, airtight space in tissue homogenate pipe.
The temperature probe of the present invention, refrigeration compressor and circulating fan realize refrigeration purpose jointly.
As shown in Figure 6, the temperature measuring head of the temperature probe of the present invention adopts digital temperature sensor.The functions such as temperature sensing, signal conversion, A/D conversion are integrated on a chip by this sensor, and its internal structure is as shown in Figure 6.Temperature transition is become the signal of telecommunication by this chip, and this signal of telecommunication initially enters small signal amplifier and is amplified；Subsequently into the A/D converter of 14；Eventually pass two-wire serial digital interface output digit signals.This sensor has the calibration register can in measurement process, and calibration factor can calibrate the signal of sensor automatically.
Embodiment 2
As shown in Figures 4 and 5, it is provided that a kind of cell homogenates pipe, when sample is single cell suspension, it is possible to adopt this cell homogenates pipe.Described cell homogenates pipe includes tube wall 1 ' and is positioned at the tube core 2 ' of tube wall, described tube wall top seal, pipe lid 3 ' is stamped in bottom, described tube core 2 ' includes stator sleeve pipe 4 ' and is arranged in the rotor 5 ' of sleeve pipe 4 ', described pipe lid 3 ' is provided with rotor connection, and rotor is connected with rotor connection, and described rotor is sealed by the joint of rotor connection with pipe lid, separating the motor below pipe and provide rotary power by rotor connection for rotor, stator sleeve pipe and tube wall are fixed on motor by bayonet socket.
In described tube core, 2 ' are divided into Archimedian screw area under control and homogenate milling zone from top to bottom, and it is the hole 6 ' of 2-3mm that the stator sleeve pipe base portion of tube core is provided with some equally distributed diameters.Described Archimedian screw area under control is built with screw rod 51 ', and homogenate milling zone is built with grinding rod 52 ', and described screw rod 51 ' and grinding rod 52 ' connect to form rotor 5 ', and bottom connects rotor connection.Archimedian screw area under control is identical with embodiment 1 with the function of homogenate milling zone.
The sample size scope of cell homogenates pipe: 20-40ml；Rotor speed: 50-1000rpm, can rotate by both forward and reverse directions；Time: 1-600 second；Temperature: temperature controllable is in 0-4 DEG C；Application program: the rule of operation and homogenate program that optimize are provided according to different subcellular structure separation requirement；Sample disposal: be aseptic, airtight space in homogenate tube.
The temprature control unit of the present embodiment is identical with embodiment 1.
Above-described embodiment only for technology design and the feature of the present invention are described, its object is to allow person skilled in the art will appreciate that present disclosure and to implement according to this, can not limit the scope of the invention with this.All modifications done according to the spirit of main technical schemes of the present invention, all should be encompassed within protection scope of the present invention.
Claims (8)
1. the homogenizer being used for separating subcellular components, cell or tissue block is prepared into the cell homogenates or tissue homogenate that comprise various subcellular components, described homogenizer includes homogenate tube, homogenate tube includes tissue homogenate pipe and cell homogenates pipe, its common trait is in that: described homogenate tube includes tube wall and is positioned at the tube core of tube wall, described tube wall top seal, pipe lid is stamped in bottom, described tube core includes stator sleeve pipe and is arranged in the rotor of sleeve pipe, described pipe covers and is provided with rotor connection, rotor is connected with rotor connection, described homogenizer also includes the motor being positioned at outside homogenate tube, described motor provides rotary power by rotor connection for rotor；
Archimedian screw area under control and homogenate milling zone it is divided into from top to bottom in the tube core of described cell homogenates pipe, described Archimedian screw area under control is built with screw rod, homogenate milling zone is built with grinding rod, described screw rod and grinding rod connect to form rotor, connecting rotor connection bottom grinding rod, the region of the stator sleeve inside pipe wall correspondence grinding rod of described homogenate milling zone is provided with grinding layer；
Archimedian screw area under control and homogenate milling zone and negative pressure rotary-cut district it is divided into from top to bottom in the tube core of described tissue homogenate pipe, described Archimedian screw area under control is built with screw rod, homogenate milling zone is built with grinding rod, negative pressure rotary-cut district is built with rotary-cut cutter head, described screw rod, grinding rod and rotary-cut cutter head are sequentially connected with composition rotor, connecting rotor connection bottom rotary-cut cutter head, the position of the stator sleeve inside pipe wall correspondence grinding rod of described homogenate milling zone is provided with grinding layer.
2. the homogenizer for separating subcellular components according to claim 1, it is characterised in that: the stator sleeve pipe base portion of described cell homogenates pipe is provided with some equally distributed holes.
3. the homogenizer for separating subcellular components according to claim 2, it is characterised in that: the diameter of described hole is 2.0-3.0mm.
4. the homogenizer for separating subcellular components according to claim 1, it is characterised in that: the part in the stator sleeve pipe correspondence negative pressure rotary-cut district of described tissue homogenate pipe is provided with some equally distributed holes.
5. the homogenizer for separating subcellular components according to claim 4, it is characterised in that: the diameter of described hole is 0.8-1.2mm.
6. the homogenizer for separating subcellular components according to claim 1, it is characterised in that: described rotor is sealed by the joint of rotor connection with pipe lid.
7. the homogenizer for separating subcellular components according to claim 1, it is characterised in that: it is additionally provided with protection thermal insulation cover outside described homogenate tube.
8. the homogenizer for separating subcellular components according to claim 7, it is characterized in that: described protection thermal insulation cover is built with temperature probe and refrigeration compressor, described temperature probe and refrigeration compressor are connected with a microprocessor respectively, and described refrigeration compressor is also associated with circulating fan；Described microprocessor is also connected to motor and a touch screen.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410772165.0A CN104498350B (en) | 2014-12-16 | 2014-12-16 | A kind of homogenizer for separating subcellular components |
| PCT/CN2015/081070 WO2016095458A1 (en) | 2014-12-16 | 2015-06-09 | Homogenizer for separating subcellular fractions |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201410772165.0A CN104498350B (en) | 2014-12-16 | 2014-12-16 | A kind of homogenizer for separating subcellular components |
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| CN104498350A CN104498350A (en) | 2015-04-08 |
| CN104498350B true CN104498350B (en) | 2016-07-06 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11963695B2 (en) | 2019-05-06 | 2024-04-23 | Tissuemill Technologies Llc | Atraumatically formed tissue compositions, devices and methods of preparation and treatment |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104498350B (en) * | 2014-12-16 | 2016-07-06 | 何向锋 | A kind of homogenizer for separating subcellular components |
| CN104568539B (en) * | 2014-12-16 | 2017-05-10 | 何向锋 | Automatic histocyte separator |
| CN107952544A (en) * | 2017-12-11 | 2018-04-24 | 中国人民解放军成都军区总医院 | Biological tissue's mill |
| JP7064191B2 (en) * | 2018-02-21 | 2022-05-10 | エイブル株式会社 | Cell dispersal method and cell disperser |
| CN109136088A (en) * | 2018-09-22 | 2019-01-04 | 太原市罗塞塔石生物技术有限公司 | A kind of ultrasonic cell disruption instrument |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| FR1018847A (en) * | 1950-03-31 | 1953-01-13 | Improvements to homogenizers or the like | |
| DE3047314A1 (en) * | 1980-12-16 | 1982-07-29 | Josef A. 7144 Asperg Blach | DEVICE FOR MIXING, DISPERSING AND HOMOGENIZING MASSES WITH AT LEAST ONE VISCOUS COMPONENT |
| CN2199035Y (en) * | 1994-10-13 | 1995-05-31 | 冶金工业部洛阳耐火材料研究院 | Stirrer for producing slurry containing fibre |
| CN201464269U (en) * | 2009-06-02 | 2010-05-12 | 北京必威安泰生物科技有限公司 | Biological tissue aseptic pulp homogenizer |
| CN203320013U (en) * | 2013-06-03 | 2013-12-04 | 中国科学院沈阳应用生态研究所 | Plant homogenizing device |
| CN203597361U (en) * | 2013-11-21 | 2014-05-21 | 昆明快乐王子食品有限公司 | Fruit grinding device |
| CN204281757U (en) * | 2014-12-16 | 2015-04-22 | 何向锋 | A kind of homogenizer for separating of subcellular components |
| CN104498350B (en) * | 2014-12-16 | 2016-07-06 | 何向锋 | A kind of homogenizer for separating subcellular components |
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2014
- 2014-12-16 CN CN201410772165.0A patent/CN104498350B/en active Active
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- 2015-06-09 WO PCT/CN2015/081070 patent/WO2016095458A1/en active Application Filing
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11963695B2 (en) | 2019-05-06 | 2024-04-23 | Tissuemill Technologies Llc | Atraumatically formed tissue compositions, devices and methods of preparation and treatment |
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| CN104498350A (en) | 2015-04-08 |
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