CN110420353A - A kind of extraction element and its extracting method of adipose-derived stromal vascular fraction - Google Patents
A kind of extraction element and its extracting method of adipose-derived stromal vascular fraction Download PDFInfo
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- CN110420353A CN110420353A CN201910688031.3A CN201910688031A CN110420353A CN 110420353 A CN110420353 A CN 110420353A CN 201910688031 A CN201910688031 A CN 201910688031A CN 110420353 A CN110420353 A CN 110420353A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
- A61L27/3633—Extracellular matrix [ECM]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
- A61L27/3687—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
- A61L27/3691—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by physical conditions of the treatment, e.g. applying a compressive force to the composition, pressure cycles, ultrasonic/sonication or microwave treatment, lyophilisation
Abstract
The invention discloses a kind of extraction element of adipose-derived stromal vascular fraction and its propose method, the extraction element is used to extract the stromal vascular fraction in adipose tissue, it can be realized the centrifugal movement of in vitro fatty agglomerate, eddying motion, shake movement, centrifugation concussion movement, vortex concussion movement, this method, which extracts stromal vascular fraction, has many advantages, such as that extraction efficiency is high, cell activity is good, does not introduce enzyme solution；The various kinds of cell ingredient contained in vascular stroma ingredient can be widely applied to the multidirectional field such as fat transfer, repair of cartilage；The viable count that the vortex used in its extracting method shakes resulting stromal vascular fraction is high, proliferative capacity is strong, and extracting method is simple, at low cost, safe and efficient, implementation easy to operate.
Description
Technical field
The present invention relates to fields of biomedicine, and in particular to a kind of extraction element of adipose-derived stromal vascular fraction and its
Extracting method.
Background technique
Adipose tissue is the Regeneration and Repair resource that soft tissue repair field one kind is important in recent years, can pass through liposuction
It easily obtains, and operation risk is low, patient is damaged small.Some clinical researches find adipose-derived stromal vascular fraction
(Stromal Vascular Fraction, SVF) has apparent anti-inflammatory and early stage vascularization, fatty group for adding SVF
The effect that transplantation effect is substantially better than simple adipose tissue is knitted, prompts SVF to have in tissue repair and regeneration field wide
Prospect.
Currently, from adipose tissue separation and Extraction SVF mode mainly pass through enzymic digestion or ultracentrifugal method into
Row.Wherein, the more demanding aseptic condition of enzyme digestion, and need to introduce exogenous biological enzyme, it takes a long time, it is with higher
Potential risk is chiefly used in preparing SVF under laboratory condition, and not yet approval is applied in clinic.And supercentrifugal process is a kind of
Mechanical preparation method, it is comparatively safe, but its SVF activity extracted is poor, and the improvement postoperative to fat transfer still needs to be mentioned
It is high.
Therefore, how under the premise of not introducing exogenous biological reagent, efficient, safety preparation high activity
SVF is always clinical urgent problem to be solved.
Summary of the invention
In order to solve the above problem, the present invention provides extraction element and its side of mentioning of a kind of adipose-derived stromal vascular fraction
Method, the extraction element are used to extract stromal vascular fraction in adipose tissue, can be realized in vitro fatty agglomerate centrifugal movement,
Eddying motion, concussion movement, movement is shaken in centrifugation, be vortexed concussion movement, and this method extraction stromal vascular fraction, which has, extracts effect
Rate is high, cell activity is good, does not introduce the advantages that enzyme solution.The various kinds of cell ingredient contained in vascular stroma ingredient can be widely applied to
The multidirectional field such as fat transfer, repair of cartilage；The vortex used in its extracting method shakes the work of resulting stromal vascular fraction
Cell number is high, proliferative capacity is strong, and extracting method is simple, at low cost, safe and efficient, implementation easy to operate.
In order to achieve the above object, the present invention is resolved using following technical scheme.
(1) a kind of extraction element of adipose-derived stromal vascular fraction characterized by comprising bottom plate, mounting plate, support
Disk, rack for test tube, first driving device, the second driving device and third driving device；Wherein, the upper end of the bottom plate is fixed with
One position-limiting tube, the bottom end of the mounting plate are fixed with the second position-limiting tube to match with the first position-limiting tube, first position-limiting tube
Sliding is inserted in second position-limiting tube；The mounting plate is driven to move up and down by first driving device on the bottom plate；Institute
The middle part upper end for stating mounting plate drives the pallet to horizontally rotate by the second driving device；The upper end of the pallet passes through third
Driving device driving rack for test tube horizontally rotates；The rack for test tube includes connecting rod and support frame, the lower end of the connecting rod and described the
The output shaft of three driving devices is fixedly connected；The upper end of the connecting rod is detachably and fixedly provided with support frame, support frame as described above last week
To multiple snap rings are symmetrically arranged with, the snap ring is for blocking test tube；The upper end bilateral symmetry of the mounting plate is fixed with first
Bracket is fixed with support roller in the first support, and the bottom end of the pallet is provided with the sliding rail to match with the support roller.
The extraction element of adipose-derived stromal vascular fraction according to the present invention, the second position-limiting tube on mounting plate can be the bottom of at
Upper and lower opposite sliding, first driving device drive installation plate vertical tremor in the first position-limiting tube on plate, and then drive and installation
The rack for test tube of plate connection shakes movement up and down.Multiple snap rings on rack for test tube are used in test tube for disposably placing multiple test tubes
In placing object to be extracted (object to be extracted in the application is in vitro fatty agglomerate), the working efficiency of object to be extracted is improved；Connecting rod
It is detachably connected with support frame, different support frames can be used instead according to actual needs.In the first support of the two sides of mounting plate
Support roller can slide on the sliding rail on pallet, and the support roller in first support is used to support pallet horizontal rotation, the second driving device
For driving pallet to rotate horizontally, realize that object to be extracted revolves around the output shaft of the second driving device.Third driving device is used
It is rotated horizontally in driving rack for test tube, realizes that object to be extracted surrounds the output shaft rotation of third driving device, and then realize to be extracted
The centrifugal movement of object.The rotation and revolution that the extraction element of adipose-derived stromal vascular fraction of the invention passes through object to be extracted
It combines, realizes the eddying motion of object to be extracted；The concussion campaign that object to be extracted is realized by first driving device, passes through third
Driving device realizes the centrifugal movement of object to be extracted.
Preferably, support frame as described above hingedly or is welded with the snap ring.
The extraction element of adipose-derived stromal vascular fraction according to the present invention, support frame of the invention are divided into two types
Type, it is hinged with snap ring that one is support frames, another support frame and the welded molding of snap ring.It is carried out when needing to treat extract
When centrifugation, support frame and the hinged support frame of snap ring are selected, under the action of third driving device, the test tube on snap ring has been got rid of
Come, the centrifugation of object to be extracted is realized according to the difference of molecular weight of material size.When need to treat extract do eddying motion when, choosing
The support frame that support frame and snap ring are welded to connect is selected to realize under the action of the second driving device and third driving device wait mention
Take the eddying motion of object.
Preferably, the top of the connecting rod has a screw hole, the center of support frame as described above has a through-hole, the screw hole and
Hand the turn of the screw is equipped in through-hole, the hand the turn of the screw passes through through-hole and is screwed into screw hole and is fixedly connected with support frame with connecting rod.
The screw rod of the extraction element of adipose-derived stromal vascular fraction according to the present invention, hand the turn of the screw passes through on support frame
The screw hole for passing through and being screwed into connecting rod in, realize being detachably connected for connecting rod and support frame, and then can change according to actual needs
It is used as the support frame of eddying motion or centrifugal movement.
Preferably, the first driving device includes first motor, cam and idler wheel；Wherein, in the bottom plate
Portion is fixedly installed with first motor by second support, and cam, the mounting plate are fixed on the output shaft of the first motor
The upper position opposite with the cam is provided with idler wheel.
The extraction element of adipose-derived stromal vascular fraction according to the present invention, first motor are fixed on by second support
On bottom plate, the output shaft band moving cam of first motor moves in a circle, and is moved up and down, is realized to be extracted by cam driven idler wheel
Concussion campaign up and down.In addition, having symmetrical two lug bosses on cam, cam, can during doing a circular motion
Make object concussion to be extracted twice, improves the extraction efficiency of object to be extracted.
Preferably, second driving device includes the second motor, second motor is fixed on the mounting plate
Middle part, the output shaft of second motor is fixedly connected with the pallet.
The extraction element of adipose-derived stromal vascular fraction according to the present invention, output shaft and the pallet of the second motor are fixed
Connection is rotated horizontally by the second motor driven pallet, and rack for test tube can do revolution fortune around the output shaft of the second motor on pallet
It is dynamic.
Preferably, the third driving device includes third motor, the third motor has mounting base, the support
The center of disk offers eccentric groove, and the pallet radially or/and is circumferentially uniformly provided with multiple volution troughs；The eccentric groove with
The volution trough is detachably connected with the mounting base respectively；The output shaft and the connecting rod of the third motor pass through shaft coupling
Connection.
The extraction element of adipose-derived stromal vascular fraction according to the present invention, the mounting base and support of the bottom of third motor
Eccentric groove or volution trough on disk are to be detachably connected, according to actual needs, the difference that third motor can be fixed on pallet
Position.When third motor is fixed on eccentric groove, starting the second motor, it can be achieved that object to be extracted centrifugal movement；Work as third
When motor is fixed on volution trough, start the second motor and third motor, it can be achieved that object to be extracted eddying motion.In addition, working as
When third motor is fixed on eccentric groove, start first motor and the second motor, it can be achieved that movement is shaken in the centrifugation of object to be extracted；
When third motor is fixed on volution trough, starting first motor, the second motor and third motor, it can be achieved that object to be extracted whirlpool
Rotation concussion movement.Pallet is radially even to offer multiple volution troughs, the second motor output shaft of adjustable third motor distance
Distance, realize that object to be extracted does eddying motion along different radii.Pallet is circumferentially uniformly provided with multiple volution troughs, can locate simultaneously
Multiple object samples to be extracted are managed, treatment effeciency is high.
Preferably, the bottom end of the mounting base has external screw thread, be respectively provided on the eccentric groove and volution trough with
The screw hole that the external screw thread matches.
The extraction element of adipose-derived stromal vascular fraction according to the present invention, mounting base pass through the external screw thread being mutually matched
It realizes with the internal screw thread on screw hole and is detachably connected with eccentric groove and volution trough, convenient for the position of adjusting third motor and to it
It is fixed.
Preferably, further including spring, the both ends of the spring are fixedly connected with bottom plate and mounting plate respectively.
The extraction element of adipose-derived stromal vascular fraction according to the present invention, the elastic reaction of spring are used to support installation
Plate reduces the pressure of cam and idler wheel.
Preferably, further including protective cover, the protective cover is used to cover entire adipose-derived stromal vascular fraction
Extraction element.
The extraction element of adipose-derived stromal vascular fraction according to the present invention, protective cover is for providing sterile and temperature control ring
Border makes object to be extracted extract or store between 4-37 DEG C, reduces the pollution that environment treats extract.
(2) a kind of extracting method of adipose-derived stromal vascular fraction, comprising the following steps:
Step 1, the tissue fascial of in vitro fatty agglomerate is rejected, shreds, obtains particulate matter；
Step 2, physiological saline is added in Xiang Suoshu particulate matter, be vortexed concussion, obtains suspension；
Step 3, the suspension is centrifuged, obtains centrifugation object；
Step 4, the centrifugation object is filtered, obtains stromal vascular fraction.
Preferably, in step 2, the frequency of the concussion that is vortexed is 5~50Hz, the time for the concussion that is vortexed is 2~
25min。
It is further preferred that the frequency of the concussion that is vortexed is 30Hz in step 2, the time for the concussion that is vortexed is 15min.
Preferably, the revolving speed of the centrifugation is 500~1500rpm in step 3, the time of centrifugation is 1~5min.
It is further preferred that the revolving speed of the centrifugation is 1000rpm in step 3, the time of centrifugation is 2min.
Detailed description of the invention
The present invention is described in further details in the following with reference to the drawings and specific embodiments.
Fig. 1 is the structural schematic diagram of the fatty agglomerate under quiescent condition；
Fig. 2 is the structural schematic diagram of the fatty agglomerate under high speed centrifugation state in the prior art；
Fig. 3 is the structural schematic diagram of the fatty agglomerate under the vortex concussion state of the application；
Fig. 4 is the structural schematic diagram of the extraction element of adipose-derived stromal vascular fraction of the invention；
Fig. 5 is the first structural schematic diagram of rack for test tube in Fig. 4；
Fig. 6 is the top view of Fig. 5；
Fig. 7 is second of structural schematic diagram of rack for test tube in Fig. 4；
Fig. 8 is the top view of the support frame in Fig. 7；
Fig. 9 is the SVF suspension Trypan Blue result extracted under different disposal method；Wherein, ordinate is viable count
(cell counts), unit are × 105；Abscissa is different processing method, and respectively low-frequency eddy shakes
(vibration), enzyme disappears (enzyme digestion), high speed centrifugation (centrifugation), nanometer emulsified (nano
emulsion)；
Figure 10 is the proliferative capacity test result of the SVF suspension extracted under different disposal method；Wherein, abscissa is number of days
(Days), ordinate is the absorbance value (Absorbance of 450OD) at 450nm；
Figure 11 is the B ultrasound testing result of the SVF suspension extracted under different disposal method；Wherein, abscissa is time, unit
For all (week)；Ordinate is volume sustainment rate (survival rate), unit %；
Figure 12 is the oil red O stain test result after the SVF suspension extracted under different disposal method is implanted into 6 months；Wherein,
Top is from left to right respectively that A- concussion group, B- enzyme disappear group in figure；Lower section is from left to right respectively C- centrifugation group, D- emulsification in figure
Group；
Figure 13 is the HE dye test result after the SVF suspension extracted under different disposal method is implanted into 6 months；Wherein, scheme
Middle top is from left to right respectively that A- concussion group, B- enzyme disappear group；Lower section is from left to right respectively C- centrifugation group, D- emulsification in figure
Group.
In the above figure: 1 bottom plate；101 first position-limiting tubes；102 second supports；2 mounting plates；201 second position-limiting tubes；202
First support；203 support rollers；3 pallets；301 eccentric grooves；302 volution troughs；4 rack for test tubes；410 connecting rods；420 support frames；421 snap rings；
422 through-holes；430 test tubes；440 hand the turn of the screws；5 first motors；501 cams；502 idler wheels；6 second motors；7 third motors；701
Mounting base；8 springs.
Specific embodiment
Fig. 1 is the structural schematic diagram of the fatty agglomerate under quiescent condition；As shown in Figure 1, fat cell and SVF cell are uniform
Distribution is in vitro.
Fig. 2 is the structural schematic diagram of the fatty agglomerate under high speed centrifugation state in the prior art；As shown in Figure 2, in high speed
Under apocarpy (centrifugal rotational speed is in 1500rpm), fat cell compressive deformation is squeezed at one piece, and SVF is struggle with from fat cell
Between pass through in narrow gap, and sink to tube bottom, and SVF is pressurized that deformation occurs, activity is affected.It follows that high
Fat cell separates more difficult with SVF cell under fast apocarpy, and the activity of the SVF cell of extraction is weaker.
Fig. 3 is the structural schematic diagram of the fatty agglomerate under the vortex concussion state of the application；From the figure 3, it may be seen that being shaken being vortexed
It swings under state, the gap between fat cell shows expanding or shrinking for rhythmicity with concussion movement, in the iuntercellular of increase
At gap, SVF cell can smoothly pass through out of gap and sink to tube bottom.It follows that be vortexed concussion be conducive to fat cell with
The separation of SVF cell, convenient for extracting SVF cell, and the activity of the SVF cell extracted is stronger.
With reference to Fig. 4-6, a kind of adipose-derived stromal vascular fraction that the embodiment of content according to the present invention is proposed
Extraction element, comprising: bottom plate 1, mounting plate 2, pallet 3, rack for test tube 4, first driving device, the second driving device and third driving
Device；Wherein, the upper end of the bottom plate 1 is fixed with the first position-limiting tube 101, and the bottom end of the mounting plate 2 is fixed with and the first limit
The second position-limiting tube 201 that position pipe 101 matches, the sliding of the first position-limiting tube 101 are inserted in second position-limiting tube 201；Institute
State drives the mounting plate 2 to move up and down on bottom plate 1 by first driving device；The middle part upper end of the mounting plate 2 passes through the
Two driving devices drive the pallet 3 to horizontally rotate；The upper end of the pallet 3 drives 4 water of rack for test tube by third driving device
Flat turn is dynamic；The rack for test tube 4 includes connecting rod 410 and support frame 420, the lower end of the connecting rod 410 and the third driving device
Output shaft be fixedly connected；The upper end of the connecting rod 410 is detachably and fixedly provided with support frame 420, circumferential direction on support frame as described above 420
Multiple snap rings 421 are symmetrically arranged with, the snap ring 421 is for blocking test tube 430；The upper end bilateral symmetry of the mounting plate 2 is solid
Surely there is first support 202, support roller 203 is fixed in the first support 202, the bottom end of the pallet 3 is provided with and the support
The sliding rail that wheel 203 matches.
In the embodiment above, the second position-limiting tube 201 on mounting plate 2 can be in the first position-limiting tube 101 on bottom plate 1
Upper and lower opposite sliding, 2 vertical tremor of first driving device drive installation plate, and then drive the rack for test tube 4 connecting with mounting plate 2
Concussion movement up and down.Multiple snap rings 421 on rack for test tube 4 are used to put for disposably placing multiple test tubes 430 in test tube 430
Object to be extracted (object to be extracted in the application is in vitro fatty agglomerate) is set, the working efficiency of object to be extracted is improved；Connecting rod 410 with
Support frame 420 is detachably connected, and can use different support frames 420 instead according to actual needs.The first support of the two sides of mounting plate 2
Support roller 203 on 202 can slide on the sliding rail on pallet 3, and it is horizontal that the support roller 203 in first support 202 is used to support pallet 3
Rotation, the second driving device realize that object to be extracted is public around the output shaft of the second driving device for driving pallet 3 to rotate horizontally
Turn.Third driving device realizes that object to be extracted surrounds the output shaft of third driving device certainly for driving rack for test tube 4 to rotate horizontally
Turn, and then realizes the centrifugal movement of object to be extracted.The extraction element of adipose-derived stromal vascular fraction of the invention passes through wait mention
The rotation of object and combining for revolution are taken, realizes the eddying motion of object to be extracted；Object to be extracted is realized by first driving device
Concussion campaign, the centrifugal movement of object to be extracted is realized by third driving device.
With reference to Fig. 5-8, according to one embodiment of present invention, support frame as described above 420 hingedly or is welded with the snap ring 421
It connects.
In the embodiment above, support frame 420 of the invention divides for two types, and one is support frames 420 and snap ring 421
Hingedly (as seen in figs. 5-6)；Another kind is support frame 420 and the welded molding (as Figure 7-8) of snap ring.When needs are treated
When extract is centrifuged, support frame 420 and the hinged support frame 420 of snap ring 421 are selected, in the effect of third driving device
Under, the test tube 430 on snap ring 421 is thrown up, and the centrifugation of object to be extracted is realized according to the difference of molecular weight of material size.When need
When treating extract and doing eddying motion, the support frame 420 for selecting support frame 420 and snap ring 421 to be welded to connect, in the second driving
Under the action of device and third driving device, the eddying motion of object to be extracted is realized.
With reference to Fig. 4-8, according to one embodiment of present invention, the top of the connecting rod 410 has screw hole, support frame as described above
420 center has through-hole 422, is equipped with hand the turn of the screw 440 in the screw hole and through-hole 422, the hand the turn of the screw 440 passes through
Support frame 420 is fixedly connected by through-hole 422 and being screwed into screw hole with connecting rod 410.
In the embodiment above, the screw rod of hand the turn of the screw 440 passes through passing through and being screwed into connecting rod 410 on support frame 420
In screw hole, realization connecting rod 410 is detachably connected with support frame 420, and then can be used instead according to actual needs and be done eddying motion
Or the support frame 420 of centrifugal movement.
With reference to Fig. 4, according to one embodiment of present invention, the first driving device includes first motor 5, cam 501
With idler wheel 502；Wherein, the middle part of the bottom plate 1 is fixedly installed with first motor 5, the first motor by second support 102
It is fixed with cam 501 on 5 output shaft, the position opposite with the cam 501 is provided with idler wheel 502 on the mounting plate 2.
In the embodiment above, first motor 5 is fixed on bottom plate 1 by second support 102, the output of first motor 5
Axis band moving cam 501 moves in a circle, and drives idler wheel 502 to move up and down by cam 501, realizes the fortune of concussion up and down to be extracted
It is dynamic.In addition, having symmetrical two lug bosses on cam 501, cam 501 can make during doing a circular motion wait mention
It takes object concussion twice, improves the extraction efficiency of object to be extracted.
With reference to Fig. 4, according to one embodiment of present invention, second driving device include the second motor 6, described second
Motor 6 is fixed on the middle part of the mounting plate 2, and the output shaft of second motor 6 is fixedly connected with the pallet 3.
In the embodiment above, the output shaft of the second motor 6 is fixedly connected with pallet 3, drives pallet by the second motor 6
3 rotate horizontally, and rack for test tube 4 can do revolution motion around the output shaft of the second motor 6 on pallet 3.
With reference to Fig. 4, according to one embodiment of present invention, the third driving device includes third motor 7, the third
Motor 7 has mounting base 701, and the center of the pallet 3 offers eccentric groove 301, and the pallet 3 is radially or/and circumferential equal
It is even to offer multiple volution troughs 302；The eccentric groove 301 detachably connects with the mounting base 701 respectively with the volution trough 302
It connects；The output shaft of the third motor 7 is connect with the connecting rod 410 by shaft coupling.
In the embodiment above, the eccentric groove 301 or volution trough in the mounting base 701 of the bottom of third motor 7 and pallet 3
302 is are detachably connected, according to actual needs, the different location that third motor 7 can be fixed on pallet 3.By third motor 7
When being fixed on eccentric groove 301, starting third motor 7, it can be achieved that object to be extracted centrifugal movement；When third motor 7 is fixed on
When on volution trough 302, start the second motor 6 and third motor 7, it can be achieved that object to be extracted eddying motion.In addition, when third electricity
When machine 7 is fixed on eccentric groove 301, start first motor 5 and the second motor 6, it can be achieved that movement is shaken in the centrifugation of object to be extracted；
When third motor 7 is fixed on volution trough 302, starting first motor 5, the second motor 6 and third motor 7 are, it can be achieved that wait mention
The vortex of object is taken to shake movement.Pallet 3 is radially even to offer multiple volution troughs 302, adjustable 7 distance of third motor
The distance of two motors, 6 output shaft realizes that object to be extracted does eddying motion along different radii.Pallet 3 is circumferentially uniformly provided with multiple
Volution trough 302, can handle multiple object samples to be extracted simultaneously, and treatment effeciency is high.
With reference to Fig. 4, according to one embodiment of present invention, the bottom end of the mounting base 701 has external screw thread, the centrifugation
The screw hole to match with the external screw thread is respectively provided on slot 301 and volution trough 302.
In the embodiment above, mounting base 701 pass through the internal screw thread on the external screw thread and screw hole that are mutually matched realize with from
Heart slot 301 and volution trough 302 are detachably connected, convenient for adjusting the position of third motor 7 and it being fixed.
Further include spring 8 according to one embodiment of present invention with reference to Fig. 4, the both ends of the spring 8 respectively with bottom plate 1
It is fixedly connected with mounting plate 2.
In the embodiment above, the elastic reaction of spring 8 is used to support mounting plate 2, reduces cam 501 and idler wheel 502
Pressure.
It according to one embodiment of present invention, further include protective cover, the protective cover is for covering entire adipose-derived base
The extraction element of matter vascular components.
In the embodiment above, protective cover is for providing sterile and controlled heat environment, make object to be extracted between 4-37 DEG C into
Row extracts or storage, reduces the pollution that environment treats extract.
Embodiment 1
A kind of extracting method of adipose-derived stromal vascular fraction, comprising the following steps:
Step 1, the tissue fascial of in vitro fatty agglomerate is rejected, shreds, obtains particulate matter.
Step 2, physiological saline is added into particulate matter, and (physiological saline uses concentration for 0.9% sodium chloride solution, uses
Amount is submergence particulate matter), and the concussion 15min that is vortexed under the conditions of frequency is 30Hz, obtain suspension.
Step 3, suspension is centrifuged 2min under 1000rpm speed conditions, obtains centrifugation object.
Step 4, centrifugation object is filtered using the strainers of 100 mesh, obtain the stromal vascular fraction of tube bottom to get.
Embodiment 2
A kind of extracting method of adipose-derived stromal vascular fraction, comprising the following steps:
Step 1, the tissue fascial of in vitro fatty agglomerate is rejected, shreds, obtains particulate matter.
Step 2, physiological saline is added into particulate matter, and (physiological saline uses concentration for 0.9% sodium chloride solution, uses
Amount is submergence particulate matter), and the concussion 2min that is vortexed under the conditions of frequency is 50Hz, obtain suspension.
Step 3, suspension is centrifuged 5min under 500rpm speed conditions, obtains centrifugation object.
Step 4, centrifugation object is filtered using the strainers of 100 mesh, obtain the stromal vascular fraction of tube bottom to get.
Embodiment 3
A kind of extracting method of adipose-derived stromal vascular fraction, comprising the following steps:
Step 1, the tissue fascial of in vitro fatty agglomerate is rejected, shreds, obtains particulate matter.
Step 2, physiological saline is added into particulate matter, and (physiological saline uses concentration for 0.9% sodium chloride solution, uses
Amount is submergence particulate matter), and the concussion 25min that is vortexed under the conditions of frequency is 5Hz, obtain suspension.
Step 3, suspension is centrifuged 1min under 1500rpm speed conditions, obtains centrifugation object.
Step 4, centrifugation object is filtered using the strainers of 100 mesh, obtain the stromal vascular fraction of tube bottom to get.
Embodiment 4
A kind of extracting method of adipose-derived stromal vascular fraction, comprising the following steps:
Step 1, the tissue fascial of in vitro fatty agglomerate is rejected, shreds, obtains particulate matter.
Step 2, physiological saline is added into particulate matter, and (physiological saline uses concentration for 0.9% sodium chloride solution, uses
Amount is submergence particulate matter), and the concussion 20min that is vortexed under the conditions of frequency is 20Hz, obtain suspension.
Step 3, suspension is centrifuged 1.5min under 1200rpm speed conditions, obtains centrifugation object.
Step 4, centrifugation object is filtered using the strainers of 100 mesh, obtain the stromal vascular fraction of tube bottom to get.
Resulting stromal vascular fraction (SVF) is configured to suspension, after then being mixed with fatty agglomerate, is obtained fatty；It will
Fat injection enters host site.Have the characteristics that be easy to vascularization, cell activity good after the stromal vascular fraction injection of preparation.Cooperation
Autologous fat transplantation postoperative body product retention ratio can be improved in autologous fat transplantation technology.Furthermore SVF can also be with a variety of drugs and material
Collocation, for treating the diseases such as osteoarthritis, repair of cartilage.Meanwhile the application is studies have shown that the concussion that is vortexed is in frequency
It is best to the extraction effect of stromal vascular fraction under the conditions of 30Hz, time are 15min.
Test 1
1) test method
Rabbit back fat tissue is aseptically extracted, is divided into quarter after rejecting fascia, low frequency whirlpool is respectively adopted
Rotation concussion (vibration), high speed centrifugation (centrifugation), enzyme disappear (enzyme digestion), are nanometer emulsified
The method of (nano emulsion) is handled, and is extracted SVF, and be configured to the SVF suspension of 1mL, is passed through Trypan Blue
(Trypan blue staining) observes the viable count at once of its preparation, and test result is as shown in Figure 9；And use CCK-
Its proliferative capacity is observed in 8 detections, and test result is as shown in Figure 10.
2) test result
As shown in Figure 9, using low-frequency eddy concussion extract SVF viable count at once be apparently higher than high speed centrifugation,
Enzyme disappears and nanometer emulsified three kinds of methods.As shown in Figure 10, CCK8 testing result shows that the SVF extracted using low-frequency eddy concussion is increased
It grows activity and is apparently higher than high speed centrifugation and nanometer emulsified method, and it is close with enzyme digestion.
Test 2
1) test method
Further, whether this advantage by vortex concussion method under zoopery verifying in vivo environment is still aobvious
It writes.24 male rabbits are divided into 4 groups, every group 6, each rabbit is tested as follows respectively: being taken rabbit back fat, is adopted
It is handled with above-mentioned four kinds of methods with the sterile cellulite of equivalent after obtaining SVF suspension, is respectively carried out with 1g fat agglomerate respectively
Mixing, and implantation in rabbit ear subcutaneous tissue of back construct rabbit ear autologous fat transplantation model.(B-ultrasound is detected using B ultrasound
Examination cellulite volume change trend) is observed, test result is as shown in figure 11；And according to group, respectively at postoperative 2
Materials are put to death when week, January, March, June, carry out oil red O stain and HE dyeing is clear at rouge effect, wherein oil red O stain is tested
As a result as shown in figure 12, HE dye test result is as shown in figure 13.
2) test result
As shown in Figure 11, B ultrasound the results show that various distinct methods to change over time trend visible postoperative remote in fat transfer
The volume sustainment rate of phase (at 6 months), vortex concussion group are apparently higher than other each groups, next is centrifugation group, enzyme disappears group and nanometer
Emulsification group.
By Figure 12 and Figure 13 it is found that vortex concussion method and centrifugal process at rouge effect more evenly, and enzyme disappear group and it is nanometer emulsified
Then visible more connective tissue is grown into group, and fat drips are smaller, are unevenly distributed.
To sum up, the SVF cell activity with higher that vortex concussion method is extracted has clinical Transformation Potential well.
Obviously, various changes and modifications can be made to the invention without departing from essence of the invention by those skilled in the art
Mind and range.In this way, if these modification and variations of the invention belong to the range of the claims in the present invention and its equivalent technologies
Within, then the present invention is also intended to include these modifications and variations.
Claims (10)
1. a kind of extraction element of adipose-derived stromal vascular fraction characterized by comprising bottom plate (1), mounting plate (2), support
Disk (3), rack for test tube (4), first driving device, the second driving device and third driving device；Wherein,
The upper end of the bottom plate (1) is fixed with the first position-limiting tube (101), and the bottom end of the mounting plate (2) is fixed with and the first limit
Second position-limiting tube is inserted in the second position-limiting tube (201) that position pipe (101) matches, the first position-limiting tube (101) sliding
(201) in；
The mounting plate (2) are driven to move up and down by first driving device on the bottom plate (1)；In the mounting plate (2)
Portion upper end drives the pallet (3) to horizontally rotate by the second driving device；The upper end of the pallet (3) is driven by third and is filled
Driving rack for test tube (4) is set to horizontally rotate；
The rack for test tube (4) includes connecting rod (410) and support frame (420), and the lower end of the connecting rod (410) and the third drive
The output shaft of device is fixedly connected；The upper end of the connecting rod (410) is detachably and fixedly provided with support frame (420), support frame as described above
(420) it is circumferentially symmetrically arranged on multiple snap rings (421), the snap ring (421) is for blocking test tube (430)；
The upper end bilateral symmetry of the mounting plate (2) is fixed with first support (202), is fixed on the first support (202)
The bottom end of support roller (203), the pallet (3) is provided with the sliding rail to match with the support roller (203).
2. the extraction element of adipose-derived stromal vascular fraction according to claim 1, which is characterized in that support frame as described above
(420) with the snap ring (421) hinged or welding.
3. the extraction element of adipose-derived stromal vascular fraction according to claim 1, which is characterized in that the connecting rod
(410) top has screw hole, and the center of support frame as described above (420) has through-hole (422), in the screw hole and through-hole (422)
Be equipped with hand the turn of the screw (440), the hand the turn of the screw (440) pass through through-hole (422) and be screwed into screw hole by support frame (420) with
Connecting rod (410) is fixedly connected.
4. the extraction element of adipose-derived stromal vascular fraction according to claim 1, which is characterized in that described first drives
Dynamic device includes first motor (5), cam (501) and idler wheel (502)；Wherein, pass through second support in the middle part of the bottom plate (1)
(102) it is fixedly installed with first motor (5), is fixed with cam (501), the installation on the output shaft of the first motor (5)
The position opposite with the cam (501) is provided with idler wheel (502) on plate (2).
5. the extraction element of adipose-derived stromal vascular fraction according to claim 1, which is characterized in that described second drives
Dynamic device includes the second motor (6), and second motor (6) is fixed on the middle part of the mounting plate (2), second motor
(6) output shaft is fixedly connected with the pallet (3).
6. the extraction element of adipose-derived stromal vascular fraction according to claim 1, which is characterized in that the third is driven
Dynamic device includes third motor (7), and the third motor (7) has mounting base (701), and the center of the pallet (3) offers
Eccentric groove (301), the pallet (3) radially or/and are circumferentially uniformly provided with multiple volution troughs (302)；The eccentric groove
(301) it is detachably connected respectively with the mounting base (701) with the volution trough (302)；The output shaft of the third motor (7)
It is connect with the connecting rod (410) by shaft coupling.
7. the extraction element of adipose-derived stromal vascular fraction according to claim 6, which is characterized in that the mounting base
(701) bottom end has external screw thread, is respectively provided with and the external screw thread phase on the eccentric groove (301) and volution trough (302)
The screw hole matched.
8. a kind of extracting method of adipose-derived stromal vascular fraction, which comprises the following steps:
Step 1, the tissue fascial of in vitro fatty agglomerate is rejected, shreds, obtains particulate matter；
Step 2, physiological saline is added in Xiang Suoshu particulate matter, be vortexed concussion, obtains suspension；
Step 3, the suspension is centrifuged, obtains centrifugation object；
Step 4, the centrifugation object is filtered, obtains stromal vascular fraction.
9. the extracting method of adipose-derived stromal vascular fraction according to claim 8, which is characterized in that in step 2, institute
The frequency for stating the concussion that is vortexed is 5~50Hz, and the time for the concussion that is vortexed is 2~25min.
10. the extracting method of adipose-derived stromal vascular fraction according to claim 8, which is characterized in that in step 3,
The revolving speed of the centrifugation is 500~1500rpm, and the time of centrifugation is 1~5min.
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