Fat stem cell separator

Mode of execution

The following, describes, with reference to the accompanying drawings, a detailed description, of a preferred embodiment of the present invention by, only, if it is considered that a detailed description of a related known function or structure may unnecessarily obscure the gist, of the present invention.

Also, the same reference numerals, are used, for portions having similar functions and functions throughout the drawings.

Also, in the entire specification, when a certain portion is connected " to another portion " includes the case, where " direct connection " is not included and, includes " some structural elements in a case " where different elements are disposed therebetween, and, may include other structural elements, " instead of excluding other structural elements, " in the case where the other components are not specifically, described.

1a And 2, the adipose-derived stem cell separation device 100 according to a preferred embodiment of the present invention may include first a cylindrical case 10, first of the cylindrical outer cap 20, second and 30, a sealing cover 40 50, so as to be able to easily separate and obtain the adipose stem cells, second from the adipose tissue by using a container.

1a, 1b And 2, the sealing cover, is provided with a lid member first for selectively opening or closing an open upper portion of the 10-cylinder case 11 by a lever 12, extending through a communication port 13 formed in an upper surface thereof so first as to 20 extend in a predetermined length from the, lower portion.

The auxiliary cover 12 may be formed of a cover member having an internal thread portion threadedly engaged with an external thread portion formed at an outer surface of the hollow tube 11 extending from the communication port 11a. at a predetermined height.

The lever 13 may be integrally provided at a lower surface first of the 10 sealing cover, and a rod member having a predetermined length moving together with the first sealing cover at an upper end thereof.

As shown 1b and 2, the joystick 13 may be formed of a rod member formed at a predetermined length of the upper end by a rod member forming first formed at a predetermined length of the upper end of the cap extension pipe 10 provided in the closed lower face 14 of the sealing cover, described above and a rod member formed integrally at a predetermined length of the closed lower surface first of the cover 10.

Although, is illustrated in such a manner that the lever 13, is integrally bonded to the closed lower surface first of, sealing cover by first sealing cover rotation operation, but not limited, the upper and lower movement first may be performed by a separate operation to the, sealing cover.

That is, as shown 3a and 3b, the lever 13 is extended 14 from the through-hole formed in the closed upper surface of, of the cap extension pipe, and the upper end portion of the lever 14 is exposed to the outside through the cover extension pipe first so that the upper portion 10 of the seal cover, first faces outwardly 10 13, respectively 14.

, Through the first sealing cover 10 or the operation lever 13 by the positive or reverse direction rotating operation, the lever 13 can move, up and down 20a in such a manner that the neck hole 21 of the neck described later can be selectively opened and closed.

1a, And 1b, the outer peripheral surface 2 of the, cylindrical case first which is threadedly engaged with the inner threaded portion 20 formed in the lower end inner peripheral surface, of the sealing cover first is selectively opened and closed by 10 . 15. The hollow cylindrical member 25, forming first space S1 first 1st 10 of a predetermined size is 20 formed on the open upper end outer peripheral surface.

, The 1st-cylinder housing 20 seals the O-type ring member 23 provided on the upper end outer surface as the medium, with the first sealing cover 10 and then performs the sealing treatment.

And, the first sealing cover 10 is provided with a silicon plate, forming a cross slit in a region corresponding to the communication port opened and closed by the auxiliary cover to prevent external air from flowing into, space, when injection of an injection object such as an injection port first of an injection port of a syringe opened through separation, of the auxiliary cover is injected.

Further, the first sealing cover 10 includes at least one one-way check valve first forming a passage discharged from S1 space, to the outside so that air, first space S1 is smoothly discharged to the outside, when injection of an object such as an adipose-derived stem cell extract is injected, thereby preventing the internal pressure first from unnecessarily increasing excessively.

1a, 1b, And 2, the 4-cylinder housing, is a hollow cylindrical member second that communicates the neck hole 30 of the neck 13 formed at the lower end of first cylindrical housing 20 with the joystick 20a as a medium 21 with, space first of 20-cylinder case first described above. S1.

The neck 20a is formed so as to extend from first cylindrical case 20 to neck hole 21 so as to gradually decrease in inner diameter, so as to extend from the neck hole 21 toward the inside diameter of second cylindrical case 30, thereby prolonging the hourglass cross-sectional shape.

The neck hole, 20a of the neck 21 includes an upper guide tube 22, extending at a predetermined height so as to guide the liquid that fills the inner space 13 of the upper guide tube, into 22. The neck hole 22a, may include a lower guide tube 21 extending at a predetermined length to the lower portion. The neck hole second 21, may include the 20 neck hole portion formed at 30 24. ° or 1st.

Also, at least one 13-type ring 21 that generates a sealing force in contact with an inner surface of the neck hole O may be included at a lower end outer surface of the joystick 13a.

, The first cylinder case 20 and second cylinder case 30 integrally connect the neck 20a as the medium, to each other, and, while the neck hole 21 communicates, with each other as a medium, to selectively open and close the neck hole 13. by the joystick moving up and down when the rotary operation is performed.

That is, if the lower end of the lever 13 is located at the neck hole 21 or the lower guide tube 24, the neck hole 21 is positioned in the closed state, and a portion of the slit is opened, in accordance with the position of the lower end of the joystick.], the liquid filled 22 space 22a, of the, cylinder housing, can be discharged to first cylinder housing 20 through the slit first and the 30 neck hole 22a second described above, respectively 21.

As shown 1a, FIGS 1b, through 2 and 4, the cover connecting portion, is detachably assembled to the open lower portion 40 of second cylindrical case 30 so as to form, space second having a volume relatively smaller than 30 of the space first in the inside of S1 cylindrical case second S2.

For such a cover connection portion 40, a discharge port, is formed in the center of the body 41, and an assembly flange second corresponding to a fitting groove formed at an outer edge of the lower end of 30 cylindrical case, may be included at the outer side edge.

Further, preferably, is formed in a cross-sectional shape second inclined downward from the center of the 30 cylindrical case 40 so as to pass through the center of the discharge port 43 formed therein so as to increase the volume 41 of, space 40 formed between the cover connection portion second and 30 S2 cylinder case. 2nd.

The engagement of, of the open lower end of second cylindrical case 30 with the cover connecting portion 40 may adjust the volume increase or decrease second of 35 space, but the present invention is not limited to, but the present invention is not limited to the above second cylindrical case, and the cover connection portion, may be formed by elastic bonding of an assembly flange formed at a lower end of second 30 second cylindrical 40 case 30 and an outer peripheral surface of the cover, connection portion 40.

, When the neck hole 24 and the slit 13 communicate with each other by the upward movement of the lever 21 of the lower guide tube 22a, a part of the liquid filled in, the cylindrical case first is guided to 20 space 21 of the second cylinder case 30 through the neck hole second to thereby fill the S2, space second S2.

Further, when the neck hole 13 is in a closed state by the downward restoring movement of the lever 21, only the liquid corresponding to the lower layer of the liquid separated by centrifugal separation is separated and filled, space second of 30 cylinder case second S2.

1a, 1b, And 2, the 4 sealing cover, is detachably assembled to the cover member second at the lower end of the lid connection portion 50 by a closure member 40 inserted into a discharge port 41 formed in the center of the main body of the lid connection portion 51 . 40.

The closing member 51 may be formed in a column shape second extending from an upper surface of the 50 sealing cover, at a predetermined height so as to enter an inner hole of a lower extension pipe 40 extending downward from the discharge port 41 of the cover connection portion 42 to a predetermined length to seal.

The connecting tube 51 having an inner threaded portion 52a formed on the outer side of the closure member 52, may be threadedly coupled 42 with an outer threaded portion 42a formed on an outer circumferential surface of the lower extension tube, and, of the connecting tube 52 extends second from the upper surface of the 50 sealing cap, by a predetermined length.

At this time, preferably, the closure member 52 at the inner center of the connecting tube 51 is formed to be relatively higher than the height 52 of the connecting tube, so that the end of the lower extension tube 42 first enters.

And, the outside edge of the second sealing cover 50 includes an outwardly curved outer skirt portion 54, so as to have at least one 40-ring member, therebetween in combination O with the lower end outside edge of the cover connecting portion 53, whereby the sealing force can be increased.

, Of the second sealing cover 50 is combined with the cap connecting portion 52 by the combination of the connecting tube 42 and the lower extension tube 40, while the discharge port, of the cap connecting portion 42 is sealed by the closing member 51 entering the inside of the lower extension tube 40 described above 41.

On the other hand, the filter portion second for removing foreign substances contained in the adipose stem cells 50 of the subject 40 discharged from the outside through the discharge port 41 of the cap connection portion (separated by) the seal cap 60. described above may be included.

As 5 and 6, the filter portion 60 may include a flow inlet: filter body 61, communicating with the discharge port second of the cover connection portion 40 separated from 41, and an end 62; of the filter tube 64, extending from a lower surface of the upper cover 62 corresponding to the flow inlet 61 to remove the foreign matter 63 .

The filter unit 60 may include a filter case 65, in which the filter body 61 is disposed inside the open lower end, of the upper cover 64, and the open upper end part, so as to discharge the fluid filtered by the filter body to have at least one outflow port 66.

The filter body 64 may include: inside filter screen 64a, inlet end fixed to end 63 of filter pipe; and filter screen 64b, having a mesh size 64a smaller than the mesh size of inner filter screen, is disposed on outer side, of the inner filter screen.

, As the outlet end of the lower end of the inner filter screen 64a and the outlet end serving as the lower end of the outer filter screen 64b are spaced, apart from each other at a predetermined pitch.

The flow inlet 62 may be formed in the form, of a hollow tube having an internal thread formed therein so as to be threadedly coupled 41 with an external thread portion 42 formed on an outer surface of the lower extension pipe 42a extending from the discharge port.

At this time, when the inflow port 62 is formed as a hollow tube having an external thread portion formed on an outer surface, a separate adaptor tube may be connected to the lower extension tube, formed with the male screw portion 42a as the medium 42.

The outflow port 66 may be formed in a hollow tube form 70 formed on an outer surface with an external screw portion 66a screwed to an internal thread portion of an injection port of the syringe, so as to be able to collect the object 60 filtered by the filter body 64 of the filter portion.

Thus, of the final separated object second remaining in 30 space 40 between second cylinder housing S2 and cover connection (described above may be rapidly separated and extracted) without contact with air in the atmosphere after the filtration 60 has been completed by filtration unit, described above and in contact with air in the atmosphere through a separate syringe 66, communicating with the outlet. 70.

The method for separating adipose-derived stem cells from the adipose-derived stem cell separator having the above-described structure extracts the final adipose stem cells, by sequentially performing the preparation step, the fat stem cell extraction step . the primary washing step, and the secondary washing step.

In the above preparation step, the prescribed amount of adipose tissue (luer lock) is extracted from the human body by using the syringe with the luer lock, and then, of the fat tissue extracted at this time is, times each time the extracted fat tissue is repeatedly extracted.

Also, When the adipose tissue extracted, was put into the other syringe for storage, shaking the syringe causes the fat tissue to precipitate, and the inside of the syringe may be layered into the blood layer, fat extraction liquid layer and the oil layer, due to specific gravity differences.

, Workers then separate only the fat extracting solution layer, extracted between the blood layer and the oil layer from the syringe by the piston operation of the syringe described above.

In the adipose-derived stem cell extraction step, the adipose stem cells, are extracted from the fat extraction liquid obtained in the above preparation step.

That is, as illustrated 7a, of first of the sealing cover 10 closes the open upper part 12( of the first cylinder case 20 to open) the neck hole, of the neck 20a, and then enters 21 space 13 of, cylinder housing through the opened connection port P1 to 11 space first into a prescribed amount of enzyme solution first such as collagenase S1 solution, and, S1. The above-mentioned collagenase is used to hydrolyze collagen or a gelatin component (collagenase) in a prescribed. (C), amount, first of the syringe.

In this case, P1 operator inserts the fat extracting solution first into first cylindrical casing's S1 space, by the piston operation of the syringe, and then, can only extract the fat extracting solution layer precipitated in the middle layer through the piston operation of the syringe to first space first S1. of the cylindrical casing through the opened communication port.

Next, the separation device 12 of the present invention that seals the communication port 11 with the auxiliary cover 100 is loaded on the oscillation incubator (shaking incubator) to perform agitation, to perform an oscillation step, for accelerating the enzymatic reaction between the fat extraction liquid and the enzyme solution.

Also, If the separation device 100 of the present invention removed from the oscillation incubator is placed in the centrifuge, and centrifugal separation is performed at a predetermined strength by centrifugal force at a predetermined strength, the specific gravity difference is sequentially layered from the lower part of, space first of 20 cylindrical shell first to the enzyme reaction liquid S1 including the cell precipitates P of adipose-derived stem cells and the enzyme reaction C.

, As illustrated 7b, causes the joystick to rise first by the rotation of the 10 closure 13 or lever, described above such that the lower end of the joystick is located at the slit 22 of the upper guide tube 22a.

In this state, if the centrifugal separation step, is performed in the centrifuge, the lowermost cell precipitate P is transferred to 22a space 21 second of the cylindrical casing through the partially open slit second and the fully opened neck hole S2, thereby filling the second space S2.

And, if it was confirmed first second barrel casing made of a transparent material visually confirmed from the outside and the boundary layer P between the cellular precipitate C layered by the neck and the enzyme reaction liquid B was positioned in the neck hole 21, at the lower end of the joystick 13, the lever 21 was lowered 13 thereby closing the opened neck hole.

Thus, the enzyme reaction liquid first flows into C space second of second cylinder jacket by cutting off S2 cylindrical outer shell, and only the enzyme reaction solution, remains in first space of first cylindrical shell except S1 space C, of P cylinder outer shell filled with cell precipitates second S2, to achieve separation. 2nd.

At this time, the operating lever 13 moving up and down in the neck hole forms a receiving portion 13b, with a lower end opening inside the main body, so that a part of the cell precipitates separated with the capacity of the inner volume larger than second space flows into the receiving portion 13b, and thus the separation efficiency, can be improved.

In this state, as shown 7c, the syringe, assembled in communication with the lower extension pipe second of the cover connection part 50 exposed to the outside by the separation from 40 of the sealing cover 42 described above extracts the cell precipitates 71 filled in second space 30 of second cylinder case S2 P.

The enzyme reaction liquid, remaining in first space 20 of first cylinder case S1, unlike this, may be discharged to outside C through opening of the auxiliary cover 12 or opening of first sealing cover 10.

Although, is shown in FIG. 7c for cleaning the cell precipitates, remaining in second space 30 of second cylindrical casing S2, P is shown in FIG. but the present invention is not limited to second but 42 may be carried out without contact with air by other syringes that pass through injection needles inserted into closure member 71 of lower extension tube, in the state of separating the, seal cover 7b as described above, as shown in FIGS. FIGS. FIGS. and, 42 through the syringe 51 second assembled, with 50 sealing caps described above in a state of assembling the above-mentioned sealing caps.

That is, the closing member 51 penetrating through the injection needle is formed into a hollow tube pillar shape 51a having a cut region, having a predetermined thickness cut in length, or a material of such a cut-off region formed as a separate pressing member, may be composed, of a rigid flexible material having an injection needle therethrough.

As 7d and 7e, in the above-mentioned one cleaning step, the cell precipitates first are washed 20 by using a washing liquid first in S1 space W1 of P cylindrical casing, and only the cell precipitates P. which have been removed by cleaning are separated.

Such a washing liquid may be used, by selecting saline mixed with a prescribed ratio with respect to the extracted cell pellet, but other washing liquid, may be selectively used without limitation.

[1st] The opening 10 of the auxiliary cover (of the sealing cover first closing the opening of)12 cylindrical housing of the newly prepared separator device sequentially puts a predetermined amount of the cell precipitates, extracted from the fat stem cell extraction step and a predetermined amount of primary washing liquid 11, for example, brine, into P space W1 of first cylinder jacket described above in this order and is filled in first 1st space S1. S1.

At this time, preferably, the primary washing liquid W1 is dosed P in accordance with the amount of the cell precipitate, to be cleaned and a predetermined ratio.

In this state, if centrifugation, is carried out by centrifugal force of a predetermined intensity in a centrifuge for a predetermined time, the cell pellet P is washed with a washing liquid W1 while being washed, to remove contaminants, separated from the enzyme reaction liquid adsorbed on the adipocyte, fat precursor cells and stem cells included in the cell precipitates.

As shown 7d, through this centrifugation, the washed cell precipitates first and primary washing liquid 20 in first space S1 of P cylindrical casing W1 are layered, sequentially from the lower part to the upper part due to the difference in specific gravity.

Further, as shown 7e, the primary washing liquid, layered in first space 20 of first cylinder housing S1 is removed W1 by injection needle aspiration of the syringe entering through the communication port 11 opened through the auxiliary cover so that only the cell precipitates, remain in first space S1 P.

The centrifugal separation step, for sufficiently separating the cell precipitates first and the primary washing liquid P in W1 spaces is preferably 21 in a state in which the neck hole 13 is closed, by the lower end of the joystick second to maintain the cleanliness, of the space.

FIGS 7f, 7g and 7h show, in the above-described secondary cleaning step, the secondary cleaning first of the primary washed cell precipitates 20 by the washing liquid first in S1 space W2 of P cylindrical casing, separates only the cell precipitates P which have been removed by cleaning and extracts, to the outside.

That is, as shown 7f, the auxiliary cover 12 is opened, through an opened communication port 11 into W2 space first of first cylindrical casing by a predetermined amount of washing liquid S1, such as saline and mixed first with the cell precipitates P remaining in, space at a predetermined ratio.

Also, if centrifugation first is performed by the rotation of, sealing cover or joystick, as shown 7g, space and first space communication second is completely opened, and, in this state is cleaned by centrifugal force of prescribed intensity in the centrifuge for a predetermined time, W2 again removes the contaminants, separated from the enzyme reaction liquid adsorbed to, adipocyte P fat precursor cells and stem cells included in the cell precipitates.

By the centrifugal separation, the washed cell precipitates first second and secondary washing liquid 20, 30 in first second space S1, S2 of P cylindrical casing W2 are layered, sequentially from the lower part to the upper part due to the difference in specific gravity.

Also, the lower end of the lever 13 is lowered to move, and, secondary washing liquid remains in first space first of, cylinder housing when the neck hole is closed, and the washed cell precipitates second remain in second space of, cylinder housing.

At this time, the inner second of the inner hole formed in the lower end of the joystick is thus filled in the inside, of the inner hole, compared to a portion of the cell sediment having a relatively large volume of cell precipitates in. space.

, Remaining in first space 20 of first cylinder case S1 is discharged to outside W2 through the opening of auxiliary cover 12 or opening of first cover 10, and the filter unit, assembled to the lower extension pipe, of the second cylinder housing 50 30 is used to filter out the cellular 40 precipitate second from 42 the outside S2 while second foreign matter contained P in the cell precipitates P is filtered out from the outside 60, P.

That is, as shown 6, is provided in which the inlet port of the upper cover of the filter unit and the lower extension pipe second extended from the 41-seal cover are screwed together to realize communication 42 with the outflow port of the filter housing, of the filter unit and the extraction syringe 65 to realize communication 70.

When the upper portion of the cylinder housing first is opened by the 10 cover 12 or the auxiliary cover first 20 space, of 22 cylindrical housing 22a of the upper guide duct, is communicated first to each other by the space 20 of the first cylinder housing S1 remaining with the washing liquid to the outside, second S2, when the pulling operation of second the extraction syringe, is, executed. P, the inner 30 of the filter portion having, the inlet connected to the discharge port is carried out.

The cellular precipitate, transferred continuously 60 to the inside of the filter portion P is continuously extracted with the full amount 64a P 64, through the extraction syringe combined with the outflow port provided, at the lower end of the filter housing to thereby end the adipose-stem cell, separation operation 64b.

The present invention described above is not limited, to those skilled in the art, and it is obvious to those skilled in the art that, may be modified and changed, without departing from the technical idea of the present invention, or the like .