LU502898B1 - Device and method for extracting adipose-derived stromal vascular fraction - Google Patents

Device and method for extracting adipose-derived stromal vascular fraction Download PDF

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Publication number
LU502898B1
LU502898B1 LU502898A LU502898A LU502898B1 LU 502898 B1 LU502898 B1 LU 502898B1 LU 502898 A LU502898 A LU 502898A LU 502898 A LU502898 A LU 502898A LU 502898 B1 LU502898 B1 LU 502898B1
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svf
motor
driving device
tray
adipose
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LU502898A
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French (fr)
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Qianxin Lv
Ye Gao
Yan Hou
Taiqiang Dai
Liang Kong
Kai Zhang
Fuwei Liu
Yunpeng Li
Dan Jin
Feng Cao
Le Wang
Bulei Cai
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Air Force Medical Univ
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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
    • C12M45/00—Means for pre-treatment of biological substances
    • C12M45/05—Means for pre-treatment of biological substances by centrifugation

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Abstract

Disclosed are a device and a method for extracting adipose-derived stromal vascular fraction (SVF); the provided device for extracting adipose-derived SVF enables centrifugal movement, vortex movement, oscillatory movement, centrifugal oscillatory movement, vortex vibration movement of isolated adipose masses, and the provided method achieves SVF extraction with high extraction efficiency and good cell activity, with no introduction of enzyme solution; the SVF contains various cellular components that can be widely used in multi-directional fields such as adipose grafting and cartilage repair; under the vortex vibration movement of the method provided in the present application, the resulting SVF has high viable cell count and high proliferation capacity. The method for extracting is safe and simple, with low requirements for cost and operation.

Description

DEVICE AND METHOD FOR EXTRACTING ADIPOSE-DERIVED HU502898
STROMAL VASCULAR FRACTION
TECHNICAL FIELD
The invention relates to the field of biomedicine, and in particular to a device and a method for extracting adipose-derived stromal vascular fraction.
BACKGROUND
Adipose tissue is an important resource for regenerative repair in the field of soft tissue repair in recent years and can be easily obtained through liposuction with low surgical risk and minimal damage to the patient. Several clinical studies have found that adipose-derived stromal vascular fraction (SVF) has significant anti-inflammatory and early vascularization effects, and the effect of grafting adipose tissue with SVF added is significantly better than that of adipose tissue alone, suggesting that SVF has a promising future in the field of tissue repair and regeneration.
Currently, the SVF is mainly extracted from adipose tissue by enzyme digestion or high-speed centrifugation. Of these methods, enzyme digestion requires highly sterile conditions and involves the introduction of exogenous biological enzymes, which is time-consuming with relatively high potential risks, making the method mostly used for the preparation of SVF under laboratory conditions and yet to be approved for clinical application; while the method of high-speed centrifugation is a relative safer mechanical method, with poor activity of extracted SVF, and less satisfactory performance of postoperative improvement of adipose grafting.
Consequently, it has been an impending clinical problem to prepare highly active SVF in an efficient and safe manner without introducing exogenous biological reagents.
SUMMARY
In view of the above problems, the present application provides a device and a method for extracting adipose-derived stromal vascular fraction (SVF); the provided device for extracting adipose-derived SVF enables centrifugal movement, vortex movement, 502898 oscillatory movement, centrifugal oscillatory movement, vortex vibration movement of isolated adipose masses, and the provided method achieves SVF extraction with high extraction efficiency and good cell activity, with no introduction of enzyme solution; the SVF contains various cellular components that can be widely used in multi-directional fields such as adipose grafting and cartilage repair; under the vortex vibration movement of the method provided in the present application, the resulting
SVF has high viable cell count and high proliferation capacity. The method for extracting is safe and simple, with low requirements for cost and operation.
In order to achieve the above objectives, the present application adopts technical scheme as follow: (1) a device for extracting adipose-derived SVF, including: a base plate, a mounting plate, a tray, a test tube rack, a first driving device, a second driving device and a third driving device; the base plate is provided with first limiting tubes fixed on an upper end of the base plate, and the mounting plate is fixedly provided second limiting tubes matched with the first limiting tubes at a bottom end of the mounting plate; each first limiting tube is slidably sleeved inside a second limiting tube; the base plate drives the mounting plate to move up and down through the first driving device; the mounting plate drives the tray to rotate horizontally through the second driving device in an upper end of a middle part of the mounting plate; the tray drives the test tube rack to rotate horizontally through the third driving device at an upper end of the tray; the test tube rack is provided with a connecting rod and a supporting frame, where the connecting rod is fixedly connected with an output shaft of the third driving device at a lower end of the connecting rod, and the connecting rod is detachably fixed with the supporting frame at an upper end of the connecting rod; the supporting frame is provided with a plurality of snap rings symmetrically in a circumferential direction, where the snap rings are used for clamping test tubes; the mounting plate is symmetrically fixed with first brackets at an upper end of the mounting plate, where the first brackets are fixedly provided with supporting wheels, and slide rails matched with the supporting wheels are arranged at a bottom end of the tray.
According to the device for extracting adipose-derived SVF, the second limiting tube 502898 on the mounting plate can slide relatively up and down inside the first limiting tube arranged on the base plate, and the first driving device drives the mounting plate to vibrate up and down, thereby driving the test tube rack connected with the mounting plate to oscillate up and down; the snap rings on the test tube rack are used for placing a plurality of test tubes at one time, and the test tubes are used for placing to-be-extracted substances (in this application, the to-be-extracted substances are isolated adipose masses), so as to improve the to-be-extracted substances in terms of the working efficiency; the connecting rod is detachably connected with the supporting frame, with different supporting frames can be adopted according to actual requirements. The supporting wheels on the first brackets on both sides of the mounting plate slide on the slide rails on the tray; the tray is supported by the supporting wheels on the first brackets to rotate horizontally, and the tray is driven by the second driving device to rotate horizontally, achieving a revolution of the to-be-extracted substances around the output shaft of the second driving device; the third driving device drives the test tube rack to rotate horizontally, and achieves a rotation of the to-be-extracted substances around the output shaft of the third driving device, thus realizing a centrifugal movement of the to-be-extracted substances accordingly; the device for extracting adipose-derived SVF enables the vortex movement of the to-be-extracted substances by combining the rotation and revolution of the to-be-extracted substances, achieves oscillatory movement of the to-be-extracted substances through the first driving device, and also realizes the centrifugal movement of the to-be-extracted substances by the third driving device.
Optionally, the supporting frames are hinged or welded with the snap rings.
According to the device for extracting adipose-derived SVF of the present application, there is two types of supporting frames, with one being supporting frame formed by hinging supporting frame with the snap ring, the other being supporting frame integrally formed by welding supporting frame with the snap ring. When the to-be-extracted substances need to be centrifuged, the supporting frame formed by hinging supporting frame with the snap ring is selected, the test tubes on the snap rings are swung under an action by the third driving motor, and the centrifugation of 502898 the to-be-extracted substances is achieved according to different molecular weights of substances; when the to-be-extracted substance needs to be subjected to vortex movement, the supporting frame integrally formed by welding supporting frame 420 with the snap ring is selected, and the vortex movement of the to-be-extracted substances is realized under the action of the second driving device and the third driving device.
Optionally, the connecting rod is provided with a screw hole on a top end of the connecting rod, the supporting frame is provided with a through hole on a center of the supporting frame, where the screw hole and the through hole are provided with a hand screw that passes through the through hole and screw into the screw hole to fixedly connect the supporting frame with the connecting rod.
According to the device for extracting adipose-derived SVF, a screw rod of the hand screw passes through the supporting frame and is screwed into the screw hole of the connecting rod, so that the connecting rod can be detachably connected with the supporting frame, and then the supporting frame which enables vortex movement or centrifugal movement can be changed according to actual requirements.
Optionally, the first driving device includes a first motor, a cam and a roller; the first motor is fixedly installed in a middle part of the base plate through a second bracket, the cam is fixed on an output shaft of the first motor, and the roller is arranged on the mounting plate at a position opposite to the cam.
According to the device for extracting adipose-derived SVF, the first motor is fixed on the base plate through the second bracket, the output shaft of the first motor drives the cam to make a circular movement, and the cam drives the roller to move up and down, thus realizing the up-and-down oscillatory movement of the to-be-extracted substances; moreover, the cam is provided with two symmetrical bulges, with which the cam can make the to-be-extracted substances to be oscillated twice during one circular movement, thus improving the extraction efficiency of the to-be-extracted substances.
Optionally, the second driving device includes a second motor fixed at the middle part 502898 of the mounting plate, and an output shaft of the second motor is fixedly connected with the tray.
According to the device for extracting adipose-derived SVF, the output shaft of the 5 second motor is fixedly connected with the tray, and the tray is driven to rotate horizontally by the second motor, so that the test tube rack on the tray can revolve around the output shaft of the second motor.
Optionally, the third driving device involves a third motor, and the third motor is provided with a mounting seat; a center of the tray is provided with a centrifugal groove, and the tray is uniformly provided with a plurality of vortex grooves along the radial direction or/and the circumferential direction; the centrifugal groove and the vortex grooves are detachably connected with the mounting seat respectively; and an output shaft of the third motor is connected with the connecting rod through a coupling.
According to the device for extracting adipose-derived SVF, the mounting seat at a bottom of the third motor is detachably connected with the centrifugal groove or one of the vortex grooves on the tray, where the third motor can be fixed at different positions on the tray according to actual requirements. When the third motor is fixed on the centrifugal groove, the centrifugal movement of the to-be-extracted substances is realized after initiating the second motor; when the third motor is fixed on one of the vortex grooves, the vortex movement of the to-be-extracted substances is realized after initiating the second motor and the third motor; moreover, a centrifugal oscillation movement of the to-be-extracted substances is achieved after initiating the first motor and the second motor the third motor is fixed on the centrifugal groove; the vortex vibration movement of the to-be-extracted substances is achieved after the first motor, the second motor and the third motor are initiated when the third motor is fixed on one of the vortex grooves. That several vortex grooves are uniformly provided on the tray along a radial direction of the tray, with distance between the third motor and the output shaft of the second motor can be adjusted to realize the vortex movement of the to-be-extracted substances along different radii; and that several vortex grooves uniformly provided on the tray along the radial direction enable a processing of a 502898 plurality of to-be-extracted substances at the same time with high processing efficiency.
Optionally, the mounting base is provided with external threads at a bottom end of the mounting base, and the centrifugal groove and the vortex grooves are respectively provided with screw holes matched with the external threads.
According to the device for extracting adipose-derived SVF, the mounting base is detachably connected with the centrifugal groove and the vortex grooves through the external threads with mutually matched internal threads arranged on the screw holes, so that the third motor can be conveniently adjusted and fixed at different positions.
Optionally, the device includes several springs, where both ends of each spring are fixedly connected with the base plate and the mounting plate respectively.
According to the device for extracting adipose-derived SVF, the springs are used for elastically supporting the mounting plate as well as reducing pressure between the cam and the roller.
Optionally, the device includes a protective cover used for covering the device for extracting adipose-derived SVF.
According to the device for extracting adipose-derived SVF, the protective cover provides a sterile and temperature-controlled environment, so that the to-be-extracted substances are extracted and stored at 4 - 37 degree Celsius (°C) while preventing pollution from the environment to the to-be-extracted substances. (2) A method for extracting adipose-derived SVF, including:
S1, removing tissue fascias of the isolated adipose masses, and cutting the isolated adipose masses to obtain particles; 582, adding normal saline into the particles, followed by vortex vibration to obtain a suspension;
S3, centrifuging the suspension to obtain a centrifuge; and
S4, filtering the centrifuge to obtain SVF.
Optionally, the vortex vibration in S2 is carried out under frequency of 5 - 50 hertz (Hz) for a duration of 2 - 25 minutes (min); and further optionally, the vortex vibration in S2 is carried out under frequency of 30 for a 7502898 duration of 15 min.
Optionally, the centrifuging in S3 is performed with a speed of 500 - 1,500 revolutions per min (rpm) with a duration of 1-5 min; and further optionally, the centrifuging in S3 is performed with a speed of 1,000 rpm for a duration of 2 min.
BRIEF DESCRIPTION OF THE FIGURES
The present application will be further explained in detail with reference to the figures and specific embodiments below.
FIG. 1 is a structural diagram of adipose mass at rest.
FIG. 2 is a schematic diagram of a structure of adipose mass in a state of high-speed centrifugation in the prior art.
FIG. 3 is a schematic structural diagram of the adipose mass in a state of vortex vibration of the present application.
FIG. 4 is a schematic diagram of a structure of a device for extracting adipose-derived stromal vascular fraction (SVF) according to the present application.
FIG. 5 is one schematic diagram of a test tube rack of the structure in FIG. 4.
FIG. 6 1s a plan view of FIG. 5.
FIG. 7 is another schematic diagram of the test tube rack of the structure in FIG. 4.
FIG. 8 is a plan view of a supporting frame in FIG. 7.
FIG. 9 is a histogram showing results of trypan blue staining of SVF suspension extracted by different treatment methods, where the histogram used number of living cells (cell counts) with an unit of x10° as ordinate, and different treatment methods, including low-frequency vortex vibration, enzyme digestion, high-speed centrifugation and nano-emulsion, as abscissa.
FIG. 10 shows results of proliferation ability testing of SVF suspension extracted by different treatment methods, where the abscissa is duration (days), and the ordinate is absorbance value (Absorbance of 450 optical density (OD)) at 450 nanometers (nm).
FIG. 11 shows the results of B-ultrasound examination detection of SVF suspension 502898 extracted by different treatment methods, where the abscissa is duration with unit of week, and the ordinate is survival rate with unit of percent (%).
FIG. 12 shows the results of oil red O staining test of SVF suspension extracted by different treatment methods after implantation for 6 months; where the top of the figure shows from left to right, A: oscillation group and B: enzyme digestion group, respectively; and the bottom of the figure shows from left to right, C: centrifugation group and D: emulsification group respectively.
FIG. 13 shows the results of hematoxylin-eosin (HE) staining test of SVF suspension extracted by different treatment methods after implantation for 6 months, where the top of the figure shows from left to right, A: oscillation group and B: enzyme digestion group, respectively; and the bottom of the figure shows from left to right, C: centrifugation group and D: emulsification group respectively.
In the above figures: 1: base plate; 101: first limiting tube; 102: second bracket; 2: mounting plate; 201: second limiting tube; 202: first bracket; 203: supporting wheel; 3: tray; 301: centrifugal groove; 302: vortex groove; 4: test tube rack; 410: connecting rod; 420: supporting frame; 421: snap ring, 422: through hole; 430 test tube; 440: hand screw; 5: first motor; 501: cam; 502: roller; 6: second motor; 7: third motor; 701: mounting seat; and 8: spring.
DESCRIPTION OF THE INVENTION
It can be seen from FIG. 1, a structural diagram of adipose mass at rest, that adipocytes and stromal vascular fraction (SVF) cells are evenly distributed in the test tubes.
FIG. 2 is a schematic diagram of a structure of adipose mass in a state of high-speed centrifugation in the prior art, from which is can be seen that the adipocytes are pressed and deformed to squeeze together under the state of high-speed centrifugation (centrifugation speed of 1,500 revolutions per minute (rpm)), with SVF cells pass through the narrow gap between adipocytes and sink to the bottom of the tube with difficulty; SVF deforms upon pressure, with activity being affected as well; therefore,
it is difficult to separate adipocytes from SVF cells under high-speed centrifugation, 502898 and the extracted SVF cells have weak activity.
FIG. 3 is a schematic structural diagram of the adipose mass in a state of vortex vibration of the present application, from which it can be seen that the gap between adipocytes expands or shrinks rhythmically with the oscillation movement in the state of vortex vibration, SVF cells pass through the gap and sink into the bottom of the tube when the intercellular gap is enlarged, indicating that vortex vibration is beneficial to the separation of adipocytes from SVF cells, and the extracted SVF cells have strong activity.
Referring to FIGS. 4 - 6, a device for extracting adipose-derived SVF according to the embodiments of the present application includes a base plate 1, a mounting plate 2, a tray 3, a test tube rack 4, a first driving device, a second driving device and a third driving device; the base plate 1 is provided with first limiting tubes 101 fixed on an upper end of the base plate 1, and the mounting plate 2 is fixedly provided second limiting tubes 201 matched with the first limiting tubes 101 at a bottom end of the mounting plate 2; each first limiting tube 101 is slidably sleeved inside a second limiting tube 201; the base plate 1 drives the mounting plate 2 to move up and down through the first driving device; the mounting plate 2 drives the tray 3 to rotate horizontally through the second driving device in an upper end of a middle part of the mounting plate; the tray 3 drives the test tube rack 4 to rotate horizontally through the third driving device at an upper end of the tray 3; the test tube rack 4 is provided with a connecting rod 410 and a supporting frame 420, where the connecting rod 410 is fixedly connected with an output shaft of the third driving device at a lower end of the connecting rod, and the connecting rod 410 is detachably fixed with the supporting frame 420 at an upper end of the connecting rod 410; the supporting frame 420 is provided with a plurality of snap rings 421 symmetrically in a circumferential direction, where the snap rings 421 are used for clamping test tubes 430; the mounting plate 2 is symmetrically fixed with first brackets 202 at an upper end of the mounting plate 2, where the first brackets 202 are fixedly provided with supporting wheels 203,
and slide rails matched with the supporting wheels 203 are arranged at a bottom end 502898 of the tray 3.
The second limiting tube 201 on the mounting plate 2 can slide relatively up and down inside the first limiting tube 101 arranged on the base plate 1, and the first driving device drives the mounting plate 2 to vibrate up and down, thereby driving the test tube rack 4 connected with the mounting plate 2 to oscillate up and down; the snap rings 421 on the test tube rack are used for placing a plurality of test tubes 430 at one time, and the test tubes 430 are used for placing to-be-extracted substances (in this application, the to-be-extracted substances are isolated adipose masses), So as to improve the to-be-extracted substances in terms of the working efficiency; the connecting rod 410 is detachably connected with the supporting frame 420, with different supporting frames 420 can be adopted according to actual requirements. The supporting wheels 203 on the first brackets 202 on both sides of the mounting plate 2 slide on the slide rails on the tray 3; the tray 3 is supported by the supporting wheels 203 on the first brackets 202 to rotate horizontally, and the tray 3 is driven by the second driving device to rotate horizontally, achieving a revolution of the to-be-extracted substances around the output shaft of the second driving device; the third driving device drives the test tube rack 4 to rotate horizontally, and achieves a rotation of the to-be-extracted substances around the output shaft of the third driving device, thus realizing a centrifugal movement of the to-be-extracted substances accordingly; the device for extracting adipose-derived SVF enables the vortex movement of the to-be-extracted substances by combining the rotation and revolution of the to-be-extracted substances, achieves oscillatory movement of the to-be-extracted substances through the first driving device, and also realizes the centrifugal movement of the to-be-extracted substances by the third driving device.
Referring to FIGS. 5 - 8, according to one embodiment of the present application, the supporting frames 420 are hinged or welded with the snap rings 421.
In the above embodiment, there is two types of supporting frames 420, with one being supporting frame 420 formed by welding supporting frame 420 with the snap ring 421 (as shown in FIGS. 5-6), the other being supporting frame 420 integrally formed by welding with the snap ring ( as can be seen from FIGS. 7-8). When the to-be-extracted 502898 substances need to be centrifuged, the supporting frame 420 formed by hinging supporting frame 420 with the snap ring 421 is selected, the test tubes 430 on the snap rings 421 are swung under an action by the third driving motor, and the centrifugation of the to-be-extracted substances is achieved according to different molecular weights of substances; when the to-be-extracted substance needs to be subjected to vortex movement, the supporting frame 420 integrally formed by welding supporting frame 420 with the snap ring 421 is selected, and the vortex movement of the to-be-extracted substances is realized under the action of the second driving device and the third driving device.
Referring to FIGS. 4 - 8, according to one embodiment of the present application, the connecting rod 410 is provided with a screw hole on a top end of the connecting rod 410, the supporting frame 420 is provided with a through hole 422 on a center of the supporting frame 420, where the screw hole and the through hole 422 are provided with a hand screw 440 that passes through the through hole 422 and screw into the screw hole to fixedly connect the supporting frame 420 with the connecting rod 410.
In the above embodiment, a screw rod of the hand screw 440 passes through the supporting frame 420 and is screwed into the screw hole of the connecting rod 410, so that the connecting rod 410 can be detachably connected with the supporting frame 420, and then the supporting frame 420 which enables vortex movement or centrifugal movement can be changed according to actual requirements
Referring to FIG. 4, according to an embodiment of the present application, the first driving device includes a first motor 5, a cam 501 and a roller 502; the first motor 5 is fixedly installed in a middle part of the base plate 1 through a second bracket 102, the cam 501 is fixed on an output shaft of the first motor 5, and the roller 502 is arranged on the mounting plate 2 at a position opposite to the cam 501.
In the above embodiment, the first motor 5 is fixed on the base plate 1 through the second bracket 102, the output shaft of the first motor 5 drives the cam 501 to make a circular movement, and the cam 501 drives the roller 502 to move up and down, thus realizing the up-and-down oscillatory movement of the to-be-extracted substances;
moreover, the cam 501 is provided with two symmetrical bulges, with which the cam 502898 501 can make the to-be-extracted substances to be oscillated twice during one circular movement, thus improving the extraction efficiency of the to-be-extracted substances.
Referring to FIG. 4, according to an embodiment of the present application, the second driving device includes a second motor 6 fixed at the middle part of the mounting plate 2, and an output shaft of the second motor 6 is fixedly connected with the tray 3.
In the above embodiment, the output shaft of the second motor 6 is fixedly connected with the tray 3, and the tray 3 is driven to rotate horizontally by the second motor 6, so that the test tube rack 4 on the tray 3 can revolve around the output shaft of the second motor 6.
Referring to FIG. 4, according to an embodiment of the present application, the third driving device involves a third motor 7, and the third motor 7 is provided with a mounting seat 701; a center of the tray 3 is provided with a centrifugal groove 301, and the tray 3 is uniformly provided with a plurality of vortex grooves 302 along the radial direction or/and the circumferential direction; the centrifugal groove 301 and the vortex grooves 302 are detachably connected with the mounting seat 701 respectively; and an output shaft of the third motor 7 is connected with the connecting rod 410 through a coupling.
In the above embodiment, the mounting seat 701 at a bottom of the third motor 7 is detachably connected with the centrifugal groove 301 or one of the vortex grooves 302 on the tray 3, where the third motor 7 can be fixed at different positions on the tray 3 according to actual requirements. When the third motor 7 is fixed on the centrifugal groove 301, the centrifugal movement of the to-be-extracted substances is realized after initiating the third motor 7; when the third motor 7 is fixed on one of the vortex grooves 302, the vortex movement of the to-be-extracted substances is realized after initiating the second motor 6 and the third motor 7; moreover, a centrifugal oscillation movement of the to-be-extracted substances is achieved after initiating the first motor 5 and the second motor 6 when the third motor 7 is fixed on the centrifugal groove 301; the vortex vibration movement of the to-be-extracted substances is achieved after the first motor 5, the second motor 6 and the third motor 7 are initiated HU502698 when the third motor 7 is fixed on one of the vortex grooves 302. That several vortex grooves 302 are uniformly provided on the tray 3 along a radial direction of the tray 3, with distance between the third motor 7 and the output shaft of the second motor 6 can be adjusted to realize the vortex movement of the to-be-extracted substances along different radii; and that several vortex grooves 302 uniformly provided on the tray 3 along the radial direction enable a processing of a plurality of to-be-extracted substances at the same time with high processing efficiency.
Referring to FIG. 4, according to an embodiment of the present application, the the mounting base 701 is provided with external threads at a bottom end of the mounting base 701, and the centrifugal groove 301 and the vortex grooves 302 are respectively provided with screw holes matched with the external threads.
In the above embodiment, the mounting base 701 is detachably connected with the centrifugal groove 301 and the vortex grooves 302 through the external threads with mutually matched internal threads arranged on the screw holes, so that the third motor 7 can be conveniently adjusted and fixed at different positions.
Referring to FIG. 4, according to an embodiment of the present application, the device includes several springs 8, where both ends of each spring 8 are fixedly connected with the base plate 1 and the mounting plate 2 respectively.
In the above embodiment, the springs 8 are used for elastically supporting the mounting plate 2 as well as reducing pressure between the cam 501 and the roller 502.
According to an embodiment of the present application, the device also includes a protective cover used for covering the device for extracting adipose-derived SVF.
In the above embodiment, the protective cover provides a sterile and temperature-controlled environment, so that the to-be-extracted substances are extracted and stored at 4 - 37 degree Celsius (°C) while preventing pollution from the environment to the to-be-extracted substances.
Embodiment 1
A method for extracting adipose-derived SVF, including:
S1, removing tissue fascias of the isolated adipose masses, and cutting the isolated 502898 adipose masses to obtain particles;
S2, adding normal saline (0.9 percent (%) sodium chloride solution with an amount of sufficient to submerge the particles) into the particles, followed by vortex vibration for 15 min at a frequency of 30 hertz (Hz) to obtain a suspension;
S3, centrifuging the suspension at 1,000 rpm for 2 min to obtain a centrifuge; and
S4, filtering the centrifuge with a 100-mesh filter screen to obtain SVF at the bottom of the tube.
Embodiment 2
A method for extracting adipose-derived SVF, including:
S1, removing tissue fascias of the isolated adipose masses, and cutting the isolated adipose masses to obtain particles;
S2, adding normal saline (0.9% sodium chloride solution with an amount of sufficient to submerge the particles) into the particles, followed by vortex vibration for 2 min at a frequency of 50 Hz to obtain a suspension;
S3, centrifuging the suspension at 500 rpm for 5 min to obtain a centrifuge; and
S4, filtering the centrifuge with a 100-mesh filter screen to obtain SVF at the bottom of the tube.
Embodiment 3
A method for extracting adipose-derived SVF, including:
S1, removing tissue fascias of the isolated adipose masses, and cutting the isolated adipose masses to obtain particles;
S2, adding normal saline (0.9% sodium chloride solution with an amount of sufficient to submerge the particles) into the particles, followed by vortex vibration for 25 min ata frequency of 5 Hz to obtain a suspension;
S3, centrifuging the suspension at 1,500 rpm for 1 min to obtain a centrifuge; and
S4, filtering the centrifuge with a 100-mesh filter screen to obtain SVF at the bottom of the tube.
Embodiment 4
A method for extracting adipose-derived SVF, including:
S1, removing tissue fascias of the isolated adipose masses, and cutting the isolated 502898 adipose masses to obtain particles;
S2, adding normal saline (0.9% sodium chloride solution with an amount of sufficient to submerge the particles) into the particles, followed by vortex vibration for 20 min ata frequency of 20 Hz to obtain a suspension;
S3, centrifuging the suspension at 1,200 rpm for 1.5 min to obtain a centrifuge; and
S4, filtering the centrifuge with a 100-mesh filter screen to obtain SVF at the bottom of the tube.
The obtained SVF is prepared into suspension, followed by mixing with adipose mass to obtain adipose; then the adipose added with SVF is injected into an implanted area.
The prepared SVF has the characteristics of easy vascularization and good cell activity after injection, and works to improve the volume retention rate after autologous fat transplantation in combining with autologous fat transplantation technology. Moreover, SVF can be combined with various drugs and materials to be used in the fields of osteoarthritis treatment, cartilage repair and treatments of other diseases. According to the present application, the vortex vibration has the best extraction effect on SVF by vortex vibration with conditions of frequency of 30 Hz and duration of 15 min.
Experiment 1 1) Test method
The adipose tissue on back of domestic rabbit is extracted under aseptic conditions, and is divided into four equal parts after removing fascias, followed by treatment of low-frequency vortex vibration, high-speed centrifugation, enzyme digestion and nano-emulsion, respectively; then SVF is extracted and prepared into 1 milliliter (mL) of SVF suspension, followed by observation through Trypan blue staining in terms of viable cell counts, with results as shown in FIG. 9; and cell counting kit-8 (CCK8) is used to detect and observe the suspension for proliferation ability, with results as shown in FIG. 10. 2) Test results
As can be seen from FIG. 9, the immediate viable cell count of SVF extracted by 502898 low-frequency vortex vibration is significantly higher than that of SVF extracted by methods of high-speed centrifugation, enzyme digestion, and nano-emulsion; and the results of CCK8 detection as shown in FIG. 10 indicate that the proliferation activity of SVF extracted by low-frequency vortex vibration is significantly higher than that extracted by high-speed centrifugation and nano-emulsion, yet the activity of SVF extracted by low-frequency vortex vibration is close to that of SVF extracted by enzyme digestion.
Experiment 2 1) Test method
Animal experiments are further conducted to verify whether this advantage of vortex vibration is still significant in vivo, where 24 male rabbits are divided into four groups with 6 rabbits in each group, and each rabbit is subjected to the following experiments: adipose tissues on back of domestic rabbits are extracted, then same amount of sterile adipose masses are subjected to the above four methods to obtain SVF suspensions, followed by mixing with 1 gram (g) of adipose mass and implantation into the subcutaneous tissue of the back of rabbits’ ears to construct the rabbit ear autograft model; the adipose masses are detected by B-ultrasound examination in terms of volume changes, with results as shown in FIG. 11; rabbits are sacrificed according to different groups after 2 weeks, 1 month, 3 months and 6 months respectively after implantation for taking samples, and the obtained samples are detected by oil red O staining and hematoxylin-eosin (HE) staining to determine the lipid-forming performance, where the results of oil red O staining is shown in FIG. 12 and results of
HE staining are shown in FIG. 13. 2) Test results
FIG. 11 shows results of B-ultrasound examination of various different methods of long-term postoperative performance in terms of trend over time, with volume maintenance rate of vortex vibration group being significantly higher than that of other groups, followed by centrifugation group, enzyme digestion group and — nano-emulsion group.
From FIG. 12 and FIG. 13, it can be seen that the lipid-forming performance of vortex 502898 vibration and centrifugal method is rather uniform, while enzyme digestion group and nano-emulsion group, on the other hand, see more connected tissue growing in, smaller lipid droplets and uneven distribution.
To sum up, SVF extracted by vortex vibration method has high cell activity and good clinical transformation potential.
Obviously, those skilled in the art can make various changes and modifications to the present application without departing from the spirit and scope of the present application. Thus, if these modifications and variations of the present application fall within the scope of the claims of the present application and their technical equivalents, the present application is also intended to include these modifications and variations.

Claims (5)

  1. CLAIMS LU502898
    I. A device for extracting adipose-derived stromal vascular fraction (SVF), characterized by comprising: a base plate, a mounting plate, a tray, a test tube rack, a first driving device, a second driving device and a third driving device; where the base plate is provided with first limiting tubes fixed on an upper end of the base plate, and the mounting plate is fixedly provided second limiting tubes matched with the first limiting tubes at a bottom end of the mounting plate; each first limiting tube is slidably sleeved inside a second limiting tube; the base plate drives the mounting plate to move up and down through the first driving device; the mounting plate drives the tray to rotate horizontally through the second driving device in an upper end of a middle part of the mounting plate; the tray drives the test tube rack to rotate horizontally through the third driving device at an upper end of the tray; the test tube rack is provided with a connecting rod and a supporting frame, where the connecting rod is fixedly connected with an output shaft of the third driving device at a lower end of the connecting rod, and the connecting rod is detachably fixed with the supporting frame at an upper end of the connecting rod; the supporting frame is provided with a plurality of snap rings symmetrically in a circumferential direction, where the snap rings are used for clamping test tubes; the supporting frames are hinged or welded with the snap rings; the mounting plate is symmetrically fixed with first brackets at an upper end of the mounting plate, where the first brackets are fixedly provided with supporting wheels, and slide rails matched with the supporting wheels are arranged at a bottom end of the tray; the third driving device involves a third motor, and the third motor is provided with a mounting seat; a center of the tray is provided with a centrifugal groove, and the tray is uniformly provided with a plurality of vortex grooves along the radial direction or/and the circumferential direction; the centrifugal groove and the vortex grooves are detachably connected with the mounting seat respectively; and an output shaft of the third motor is connected with the connecting rod through a coupling; and the mounting base is provided with external threads at a bottom end of the mounting 502898 base, and the centrifugal groove and the vortex grooves are respectively provided with screw holes matched with the external threads.
  2. 2. The device of claim 1, characterized in that the connecting rod is provided with a screw hole on a top end of the connecting rod, the supporting frame is provided with a through hole on a center of the supporting frame, where the screw hole and the through hole are provided with a hand screw that passes through the through hole and screw into the screw hole to fixedly connect the supporting frame with the connecting rod.
  3. 3. The device of claim 1, characterized in that the first driving device comprises a first motor, a cam and a roller; the first motor is fixedly installed in a middle part of the base plate through a second bracket, the cam is fixed on an output shaft of the first motor, and the roller is arranged on the mounting plate at a position opposite to the cam.
  4. 4. The device of claim 1, characterized in that the second driving device comprises a second motor fixed at the middle part of the mounting plate, and an output shaft of the second motor is fixedly connected with the tray.
  5. 5. A method for extracting adipose-derived SVF based on the device according to any one of claims 1 to 4, characterized by comprising: S1, removing tissue fascias of the isolated adipose masses, and cutting the isolated adipose masses to obtain particles; S2, adding normal saline into the particles, followed by vortex vibration to obtain a suspension; where the vortex vibration in S2 is carried out under frequency of 5 - 50 hertz (Hz) for a duration of 2 - 25 minutes (min); S3, centrifuging the suspension to obtain a centrifuge; where the centrifuging in S3 is performed with a speed of 500 - 1,500 revolutions per min (rpm) with a duration of 1 - 5 min; and S4, filtering the centrifuge to obtain SVF.
LU502898A 2022-10-13 2022-10-13 Device and method for extracting adipose-derived stromal vascular fraction LU502898B1 (en)

Priority Applications (1)

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LU502898A LU502898B1 (en) 2022-10-13 2022-10-13 Device and method for extracting adipose-derived stromal vascular fraction

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
LU502898A LU502898B1 (en) 2022-10-13 2022-10-13 Device and method for extracting adipose-derived stromal vascular fraction

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LU502898B1 true LU502898B1 (en) 2023-04-18

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