CN115322875A - Stem cell pretreatment device and method - Google Patents
Stem cell pretreatment device and method Download PDFInfo
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Abstract
The invention relates to cell processing and discloses a stem cell pretreatment device and a method, comprising a device main body; a pulverization pretreatment mechanism for pulverizing the tissue sample rich in stem cells; a filtering and elutriating pretreatment mechanism for elutriating the crushed stem cells; and (4) a centrifugal pretreatment mechanism for performing centrifugal treatment on the stem cells. The stem cells obtained after elutriation immediately enter a centrifugal blanking dish above a centrifugal pretreatment mechanism, filtration is carried out through a centrifugal blanking hole above the centrifugal blanking dish, the stem cells immediately enter a centrifugal cavity, the centrifugal fan in the mounting base is started, the centrifugal fan immediately drives the stem cells in the centrifugal cavity to rotate for high-speed centrifugation, meanwhile, a rotating track also keeps synchronous rotation, a vibration block connected to the inner side of the rotating track does not intermittently rub the inner wall of the centrifugal cavity, resonance occurs on the inner wall of the centrifugal cavity, and high-speed centrifugation of the centrifugal fan is matched to realize high-efficiency centrifugation of the stem cells and realize pretreatment of the stem cells.
Description
Technical Field
The invention belongs to the technical field of cell culture, and particularly relates to a stem cell pretreatment device and a stem cell pretreatment method.
Background
Mesenchymal stem cells are a very characteristic class of adult stem cells. Such cells have the potential to differentiate into a variety of mature cells, including: adipocytes, chondrocytes, osteocytes, tenocytes, and myocytes. These properties, and their developmental plasticity, have attracted considerable attention from the scientific community as they may be used in regenerative medicine research.
In the traditional culture of the umbilical cord stem cells, the human umbilical cord tissues are generally pretreated to obtain relatively pure human umbilical cord mesenchymal stem cells, and then a culture dish is adopted for culture. Enrichment of stem cells has been performed in several ways. Typical methods include patch methods, enzymatic methods, density gradient methods, elutriation methods, centrifugation, hypotonic shock induced erythrolysis, and various combinations of these methods. For example, purification of stem cells from bone marrow requires removal of red blood cells and granulocytes, which is often accomplished by density gradient centrifugation. Each of these methods has disadvantages, one of which is that a laborious washing step is required after the enrichment step is performed, typically by washing the cells by repeated methods. This step typically involves placing the enriched cell suspension into a centrifuge tube and using a centrifuge to pellet the cells to the bottom of the tube. The tube was removed from the centrifuge and the supernatant decanted from the pelleted cells. The wash solution was added to the tube and the cell pellet resuspended. These steps are typically repeated 2 to 4 times. The use process is loaded down with trivial details, and often accomplishes through the cooperation of multiunit mechanism, can't realize through a set of device.
Therefore, in response to these problems, we propose a stem cell pretreatment device and method to solve.
Disclosure of Invention
The invention aims to: aiming at the defects of the prior art, the invention provides a stem cell pretreatment device and a stem cell pretreatment method, and solves the problems that the existing device is complicated in use process, is often completed by matching a plurality of groups of mechanisms, and cannot be realized by one group of devices.
The technical scheme is as follows: in order to achieve the purpose, the invention provides the following technical scheme:
a stem cell pretreatment device comprising:
a device main body;
a pulverization pretreatment mechanism for pulverizing a tissue sample rich in (containing) stem cells;
a filtering and elutriating pretreatment mechanism for elutriating the crushed stem cell tissue sample;
a centrifugal pretreatment mechanism for performing centrifugal treatment on the stem cells;
and an upper and lower separation mechanism for layering the stem cells after centrifugation.
Further preferred technical scheme, crushing pretreatment mechanism includes:
the crushing mechanism cylinder is a cylindrical cylinder;
the crushing blanking utensil is concave and is arranged at the top end of the crushing mechanism barrel;
and the crushing blanking hole is formed in the outer surface of the crushing blanking vessel.
The crushing pretreatment mechanism further comprises:
crushing the axis;
the crushing blade is arranged on the outer surface of the crushing shaft center in a surrounding manner, rotates along with the rotation of the crushing shaft center and is arranged inside the barrel of the crushing mechanism.
Place stem cell tissue sample on smashing the unloading ware earlier, seted up on the surface of smashing the unloading ware and smashed the unloading hole, the stem cell tissue sample leaks down from smashing the unloading hole, enter into the inner chamber of rubbing crusher structure barrel, be provided with crushing blade in the inner chamber, it is rotatory that crushing blade rotates through the rotation of smashing the axle center, high-speed rotation through crushing blade, carry out preliminary crushing to stem cell tissue sample, stem cell tissue sample after smashing enters into the filtration elutriation pretreatment mechanism.
In a further preferred technical scheme, the filtering and elutriating pretreatment mechanism comprises:
the elutriation mechanism barrel is a cylindrical barrel and is connected below the crushing mechanism barrel;
the elutriation blanking utensil is concave, is arranged at the top end of the elutriation mechanism barrel and is positioned below the crushing mechanism barrel;
and the elutriation blanking hole is formed in the outer surface of the elutriation blanking vessel, and the aperture of the elutriation blanking hole is smaller than that of the crushing blanking hole.
The filtration elutriation pretreatment mechanism further comprises:
the elutriation axis is a main driving source of the filtering elutriation pretreatment mechanism;
the connecting rod is connected to the outer side of the elutriation axis;
the elutriation plate is connected to the top end of the connecting rod and is a rectangular plate;
an elutriation chamber, a cavity formed between adjacent elutriation plates.
The crushed stem cell tissue sample immediately reaches an elutriation blanking dish above a filtering and elutriation pretreatment mechanism, primary filtration is carried out through an elutriation blanking hole, the stem cell tissue sample is prevented from being blocked by blanking at the same time, the stem cell tissue sample enters a barrel of the elutriation mechanism and rotates through an elutriation axis, the elutriation axis immediately drives a connecting rod to rotate, the connecting rod immediately drives an elutriation plate to rotate, the stem cells are efficiently elutriated through the rotation of the elutriation plate, and the stem cells obtained after elutriation flow out of an elutriation cavity, so that the aim of elutriating the stem cells is fulfilled.
In a further preferred embodiment, the centrifugal pretreatment mechanism comprises:
the centrifugal mechanism barrel is a cylindrical barrel and is connected below the elutriation mechanism barrel;
the centrifugal blanking utensil is concave, is arranged at the top end of the centrifugal mechanism cylinder and is positioned below the elutriation mechanism cylinder;
and the centrifugal blanking hole is formed in the outer surface of the centrifugal blanking vessel, and the aperture of the centrifugal blanking hole is smaller than that of the elutriation blanking hole.
The centrifugal pretreatment mechanism further comprises:
a mounting base mounted at the bottom end of the centrifugal mechanism cylinder
A centrifugal fan installed inside the installation base;
a centrifugal chamber;
the rotating track is wound on the outer surface of the centrifugal cavity in a circumferential manner;
and the vibration block is attached to the inner side of the rotating track and is in lap joint with the centrifugal cavity.
The diameter of the holes of the crushing blanking holes is larger than that of the elutriation blanking holes, and the diameter of the holes of the elutriation blanking holes is larger than that of the holes of the centrifugal blanking holes.
The stem cells obtained after elutriation immediately enter a centrifugal blanking dish above a centrifugal pretreatment mechanism, filtration is carried out through a centrifugal blanking hole above the centrifugal blanking dish, the stem cells immediately enter a centrifugal cavity, the centrifugal fan in the mounting base is started, the centrifugal fan immediately drives the stem cells in the centrifugal cavity to rotate for high-speed centrifugation, meanwhile, the rotating track also keeps synchronous rotation, and a vibration block connected to the inner side of the rotating track does not stop rubbing against the inner wall of the centrifugal cavity, so that resonance occurs on the inner wall of the centrifugal cavity, and the high-speed centrifugation of the centrifugal fan is matched to realize high-efficiency centrifugation of the stem cells.
In a further preferable technical scheme, the upper and lower separation mechanisms are arranged inside the centrifugal pretreatment mechanism and vertically penetrate through the inside of the centrifugal pretreatment mechanism;
the upper and lower separating mechanism includes:
the rotating bearing is arranged at the bottom end of the upper and lower separating mechanism;
the lower layer separation structure is connected with the rotating bearing;
the connecting strip is connected above the lower-layer separation structure and is installed in an arc shape;
the mounting table is connected with the connecting strip and arranged at the top end of the connecting strip;
a high speed separation shaft;
the spiral surrounding layer is wound on the outer surface of the high-speed separation shaft and is spirally wound and installed;
an adsorption separation plate arranged on the spiral surrounding layer;
the lower layer separation structure is cellular, and the lower layer separation structure is made of sponge materials. The surface of the adsorption separation plate is coated with clear oil and does not absorb water.
Adding physiological saline with the mass solubility of 0.9% equal to the volume of the physiological saline, matching with a centrifugal pretreatment mechanism for centrifugal rotation, synchronously working an upper separation mechanism and a lower separation mechanism, rotating a rotating bearing to synchronously drive a lower separation structure to rotate, synchronously keeping a high-speed separation shaft and a spiral surrounding layer to rotate at the same time, arranging an adsorption separation plate on the outer surface of the spiral surrounding layer, enabling the lower separation structure to be honeycomb-shaped, enabling the lower separation structure to be sponge-shaped, and synchronously adsorbing the saline in a centrifugal cavity by the sponge-shaped material of the lower separation structure while the lower separation structure rotates; at the moment, the surface of the adsorption separation plate is coated with clear oil and does not absorb water, and the cells centrifuged by the centrifugal pretreatment mechanism are adsorbed by the spiral arrangement of the spiral surrounding layer, so that the separation and collection of the centrifugal cells are realized, and the pretreatment of the stem cells is realized.
In a further preferred embodiment, the stem cell pretreatment method comprises the following steps:
s1: comminuting the tissue sample enriched for dry (containing) cells;
s2: stem cell elutriation;
s3: the stem cells were centrifuged.
And S1: comminuting the tissue sample enriched for stem cells; s2: stem cell elutriation; s3: the stem cell centrifugation specifically comprises the following steps:
s1: the method for crushing the tissue sample rich in the stem cells comprises the following steps:
when the device is used, a tissue sample containing stem cells is placed on a crushing blanking dish, a crushing blanking hole is formed in the outer surface of the crushing blanking dish, the tissue sample containing the stem cells leaks from the crushing blanking hole and enters an inner cavity of a barrel of a crushing mechanism, a crushing blade is arranged in the inner cavity and rotates through the rotation of a crushing shaft center, the tissue sample containing the stem cells is primarily crushed through the high-speed rotation of the crushing blade, and the crushed tissue sample containing the stem cells enters a filtering and elutriating pretreatment mechanism;
s2: the stem cell elutriation comprises the following steps:
the crushed stem cell tissue sample immediately reaches an elutriation blanking dish above a filtering and elutriation pretreatment mechanism, and is subjected to preliminary filtration through an elutriation blanking hole, so that the stem cell tissue sample is prevented from being blocked during blanking, the stem cell tissue sample enters a barrel of the elutriation mechanism and rotates through an elutriation axis, the elutriation axis immediately drives a connecting rod to rotate, the connecting rod immediately drives an elutriation plate to rotate, the stem cell tissue sample is efficiently elutriated through the rotation of the elutriation plate, and the elutriated stem cell flows out of an elutriation cavity, so that the purpose of elutriating the stem cell is realized;
s3: the stem cell centrifugation comprises the following steps:
the stem cells after elutriation immediately enter a centrifugal blanking dish above a centrifugal pretreatment mechanism, filtration is carried out through a centrifugal blanking hole above the centrifugal blanking dish, the stem cells immediately enter a centrifugal cavity, the centrifugal fan in the mounting base is started, the centrifugal fan immediately drives the stem cells in the centrifugal cavity to rotate, high-speed centrifugation is carried out, meanwhile, a rotating track also keeps synchronous rotation, a vibration block connected to the inner side of the rotating track does not intermittently rub the inner wall of the centrifugal cavity, resonance occurs on the inner wall of the centrifugal cavity, and high-speed centrifugation of the centrifugal fan is matched to realize high-efficiency centrifugation of the stem cells;
adding physiological saline with the mass solubility of 0.9% equal to the volume of the physiological saline, matching with a centrifugal pretreatment mechanism for centrifugal rotation, synchronously working an upper separation mechanism and a lower separation mechanism, rotating a rotating bearing to synchronously drive a lower separation structure to rotate, synchronously keeping a high-speed separation shaft and a spiral surrounding layer to rotate at the same time, arranging an adsorption separation plate on the outer surface of the spiral surrounding layer, enabling the lower separation structure to be honeycomb-shaped, enabling the lower separation structure to be sponge-shaped, and synchronously adsorbing the saline in a centrifugal cavity by the sponge-shaped material of the lower separation structure while the lower separation structure rotates; at the moment, clear oil is coated on the surface of the adsorption separation plate, water is not absorbed, and the cells centrifuged by the centrifugal pretreatment mechanism are adsorbed by the spiral arrangement of the spiral surrounding layer, so that the separation and collection of the centrifuged cells are realized, and the pretreatment of the stem cells is realized.
Compared with the prior art, the technical scheme of the invention has the following beneficial effects:
1. the stem cell tissue sample pretreatment device and the stem cell tissue sample pretreatment method are characterized in that the stem cell tissue sample is placed on a crushing blanking vessel firstly, the crushing blanking hole is formed in the outer surface of the crushing blanking vessel, the stem cell tissue sample leaks from the crushing blanking hole and enters an inner cavity of a barrel of a crushing mechanism, a crushing blade is arranged in the inner cavity, the crushing blade rotates through rotation of a crushing shaft center, the stem cell tissue sample is preliminarily crushed through high-speed rotation of the crushing blade, and the purpose of crushing the stem cell tissue sample is achieved.
2. According to the stem cell pretreatment device and the stem cell pretreatment method, preliminary filtration is carried out through the elutriation blanking hole, the stem cell tissue sample is prevented from being blocked due to simultaneous blanking, the stem cell tissue sample enters the barrel of the elutriation mechanism and rotates through the elutriation axis, the elutriation axis immediately drives the connecting rod to rotate, the connecting rod immediately drives the elutriation plate to rotate, the stem cells are efficiently elutriated through rotation of the elutriation plate, and the obtained stem cells after elutriation flow out of the elutriation cavity, so that the purpose of elutriation of the stem cells is achieved.
3. The stem cells obtained after elutriation immediately enter a centrifugal blanking dish above a centrifugal pretreatment mechanism, are filtered through a centrifugal blanking hole above the centrifugal blanking dish, immediately enter a centrifugal cavity, immediately drive the stem cells in the centrifugal cavity to rotate by starting a centrifugal fan in an installation base, and perform high-speed centrifugation, meanwhile, a rotating track also keeps synchronous rotation, a vibration block connected to the inner side of the rotating track does not intermittently rub the inner wall of the centrifugal cavity, so that the inner wall of the centrifugal cavity resonates, and the high-speed centrifugation of the stem cells is realized by matching with the high-speed centrifugation of the centrifugal fan.
4. According to the stem cell pretreatment device and method, during centrifugation, physiological saline with the mass solubility of 0.9% equal to that of the stem cell is added, the stem cell is matched with a centrifugal pretreatment mechanism to rotate centrifugally, an adsorption separation plate is arranged on the outer surface of a spiral surrounding layer, a lower layer separation structure is cellular, the lower layer separation structure is made of sponge materials, and the sponge materials of the lower layer separation structure synchronously adsorb the saline in a centrifugal cavity while the lower layer separation structure rotates; at the moment, clear oil is coated on the surface of the adsorption separation plate, water is not absorbed, and the cells centrifuged by the centrifugal pretreatment mechanism are adsorbed by the spiral arrangement of the spiral surrounding layer, so that the separation and collection of the centrifuged cells are realized, and the pretreatment of the stem cells is realized.
Drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the principles of the invention and not to limit the invention. In the drawings:
FIG. 1 is a schematic view of the overall structure of the stem cell pretreatment device of the present invention.
FIG. 2 is a schematic view of the stem cell pretreatment device of the present invention in a disassembled state as shown in FIG. 1.
FIG. 3 is a perspective view of the stem cell pretreatment device of the present invention shown in FIG. 2.
FIG. 4 is a schematic view of the connection structure of the pulverizing pretreatment mechanism of the stem cell pretreatment device of the present invention.
FIG. 5 is a schematic diagram of the connection structure of the filtration and elutriation pretreatment mechanism of the stem cell pretreatment device of the present invention.
FIG. 6 is a schematic view showing the connection structure of the elutriation plate of the stem cell pretreatment device of the present invention.
FIG. 7 is a schematic view of the connection structure of the centrifugal pretreatment mechanism of the stem cell pretreatment device of the present invention.
FIG. 8 is a schematic view of the connection structure of the centrifugal chamber of the stem cell pretreatment device of the present invention.
FIG. 9 is a schematic view showing a state in which the stem cell pretreatment apparatus of the present invention is disassembled from the structure of FIG. 8.
FIG. 10 is a schematic sectional view of the stem cell pretreatment device of FIG. 9 according to the present invention.
FIG. 11 is a schematic view showing the connection structure of the upper and lower separation mechanisms of the stem cell pretreatment device of the present invention.
FIG. 12 is an enlarged view of the structure of the area A in FIG. 11 of the stem cell pretreatment device according to the present invention.
In the figure: 1. a device main body;
2. a crushing pretreatment mechanism; 21. a crushing mechanism cylinder; 22. crushing a blanking vessel; 23. crushing the blanking holes; 24. crushing the axis; 25. and (4) crushing blades.
3. A filtering and elutriating pretreatment mechanism; 31. a barrel body of the elutriation mechanism; 32. washing a blanking vessel; 33. elutriating the blanking hole; 34. elutriating the axes; 35. a connecting rod; 36. washing the plate; 37. the cavity is elutriated.
4. A centrifugal pretreatment mechanism; 41. a centrifugal mechanism cylinder; 42. a centrifugal blanking vessel; 43. centrifuging a blanking hole; 44. mounting a base; 45. a centrifugal fan; 46. a centrifugal chamber; 47. rotating the rail; 48. and a vibration block.
5. An up-down separating mechanism; 51. rotating the bearing; 52. a lower layer separation structure; 53. a connecting strip; 54. a mounting table; 55. a high speed separation shaft; 56. a helically wound layer; 57. adsorbing the partition plate.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
As shown in fig. 1 to 3, in one embodiment, a stem cell pretreatment device includes:
an apparatus main body 1;
a crushing pretreatment mechanism 2 for crushing the stem cells;
a filtration and elutriation pretreatment mechanism 3 for elutriating the crushed stem cells;
a centrifugal pretreatment mechanism 4 for performing centrifugal treatment on the stem cells;
and an upper and lower separation mechanism 5 for layering the stem cells after the centrifugation.
As shown in fig. 4, in one embodiment, a stem cell pretreatment apparatus includes:
an apparatus main body 1;
a crushing pretreatment mechanism 2 for crushing the stem cells;
a filtration elutriation pretreatment mechanism 3 for elutriating the crushed stem cells;
a centrifugal pretreatment mechanism 4 for performing centrifugal treatment on the stem cells;
an upper and lower separation mechanism 5 for layering the stem cells after the centrifugation;
the pulverization pre-treatment mechanism 2 comprises:
a crushing mechanism cylinder 21 which is a cylindrical cylinder;
a crushing blanking dish 22 which is concave and is arranged at the top end of the crushing mechanism cylinder 21;
and a crushing blanking hole 23 formed on the outer surface of the crushing blanking tray 22.
The pulverization pretreatment mechanism 2 further comprises:
a crushing shaft core 24;
the crushing blades 25 are provided on the outer surface of the crushing shaft center 24 in a surrounding manner, rotate with the rotation of the crushing shaft center 24, and are provided inside the crushing mechanism cylinder 21.
Place the stem cell on smashing unloading ware 22 earlier, smash and seted up on unloading ware 22's the surface and smashed unloading hole 23, the stem cell leaks down from smashing unloading hole 23, enter into the inner chamber of rubbing crusher structure barrel 21, be provided with crushing blade 25 in the inner chamber, crushing blade 25 rotates through the rotation of smashing axle center 24, high-speed rotation through crushing blade 25, carry out preliminary crushing to the stem cell, the stem cell after smashing enters into and filters in the elutriation preprocessing mechanism 3.
As shown in fig. 5 to 6, in one embodiment, a stem cell pretreatment device includes:
an apparatus main body 1;
a crushing pretreatment mechanism 2 for crushing the tissue sample rich in the stem cells;
a filtration and elutriation pretreatment mechanism 3 for elutriating the crushed stem cell tissue sample;
a centrifugal pretreatment mechanism 4 for performing centrifugal treatment on the stem cells;
an upper and lower separation mechanism 5 for layering the stem cells after centrifugation;
the filtration and elutriation pretreatment mechanism 3 comprises:
the elutriation mechanism cylinder 31 is a cylindrical cylinder and is connected below the crushing mechanism cylinder 21;
a washing material tray 32 which is concave, is arranged at the top end of the washing mechanism cylinder 31 and is positioned below the crushing mechanism cylinder 21;
and the elutriation blanking hole 33 is formed in the outer surface of the elutriation blanking dish 32, and the aperture of the elutriation blanking hole 33 is smaller than that of the crushing blanking hole 23.
The filtration elutriation pretreatment mechanism 3 further comprises:
an elutriation axis 34 which is a main driving source of the filtration and elutriation pretreatment mechanism 3;
a connecting rod 35 connected to the outside of the elutriation axis 34;
a elutriation plate 36 which is connected to the top end of the connecting rod 35 and is a rectangular plate;
a elutriation chamber 37, a cavity formed between said adjacent elutriation plates 36.
The crushed stem cell tissue sample immediately reaches an elutriation blanking dish 32 above a filtering and elutriation pretreatment mechanism 3, primary filtration is carried out through an elutriation blanking hole 33, blockage of stem cells during blanking is also prevented, the stem cells enter a barrel 31 of the elutriation mechanism, an elutriation shaft center 34 rotates, the elutriation shaft center 34 immediately drives a connecting rod 35 to rotate, the connecting rod 35 immediately drives an elutriation plate 36 to rotate, the stem cells are efficiently elutriated through rotation of the elutriation plate 36, and the elutriated stem cells flow out of an elutriation cavity 37, so that the aim of elutriating the stem cells is fulfilled.
As shown in fig. 7 to 10, in one embodiment, a stem cell pretreatment device includes:
an apparatus main body 1;
a crushing pretreatment mechanism 2 for crushing the stem cell tissue sample;
a filtration and elutriation pretreatment mechanism 3 for elutriating the crushed stem cells;
a centrifugal pretreatment mechanism 4 for performing centrifugal treatment on the stem cells;
the centrifugal pretreatment mechanism 4 comprises:
an upper and lower separation mechanism 5 for layering the stem cells after the centrifugation;
the centrifugal mechanism cylinder 41 is a cylindrical cylinder and is connected below the elutriation mechanism cylinder 31;
the centrifugal blanking vessel 42 is concave, is arranged at the top end of the centrifugal mechanism cylinder 41 and is positioned below the elutriation mechanism cylinder 31;
and the centrifugal blanking hole 43 is formed in the outer surface of the centrifugal blanking vessel 42, and the aperture of the centrifugal blanking hole 43 is smaller than that of the elutriation blanking hole 33.
The centrifugal pretreatment mechanism 4 further comprises:
a mounting base 44 mounted at the bottom end of the centrifugal mechanism cylinder 41;
a centrifugal fan 45 installed inside the mounting base 44;
the centrifugal chamber 46;
a rotating track 47 circumferentially wound around the outer surface of the centrifugal chamber 46;
and the vibration block 48 is attached to the inner side of the rotating track 47 and is overlapped with the centrifugal cavity 46.
The diameter of the holes of the crushing blanking holes 23 is larger than the diameter of the holes of the elutriation blanking holes 33, and the diameter of the holes of the elutriation blanking holes 33 is larger than the diameter of the holes of the centrifugal blanking holes 43.
The stem cells after elutriation immediately enter a centrifugal blanking dish 42 above a centrifugal pretreatment mechanism 4, the stem cells are filtered through a centrifugal blanking hole 43 above the centrifugal blanking dish 42, the stem cells immediately enter a centrifugal cavity 46, the centrifugal fan 45 in a mounting base 44 is started, the centrifugal fan 45 immediately drives the stem cells in the centrifugal cavity 46 to rotate for high-speed centrifugation, meanwhile, a rotating rail 47 also keeps synchronous rotation, a vibrating block 48 connected to the inner side of the rotating rail 47 continuously rubs the inner wall of the centrifugal cavity 46, resonance occurs on the inner wall of the centrifugal cavity 46, and high-efficiency centrifugation of the stem cells is realized by matching with high-speed centrifugation of the centrifugal fan 45.
As shown in fig. 11 to 12, in one embodiment, the upper and lower separation mechanisms 5 are arranged inside the centrifugal pretreatment mechanism 4, and penetrate through the inside of the centrifugal pretreatment mechanism 4 up and down;
the up-down separating mechanism 5 includes:
a rotary bearing 51 provided at the bottom end of the vertical separating mechanism 5;
a lower separation structure 52 connected to the rotary bearing 51;
a connecting bar 53 connected above the lower separation structure 52 and installed in an arc shape;
a mounting table 54 connected to the connecting bar 53 and provided at the top end of the connecting bar 53;
a high-speed separation shaft 55;
a spiral winding layer 56 wound around the outer surface of the high-speed separation shaft 55 and spirally wound;
and an adsorption separation plate 57 disposed on the spiral wrap layer 56.
The lower layer separation structure 52 is honeycomb-shaped, and the lower layer separation structure 52 is made of sponge. The surface of the adsorption separation plate 57 is coated with clear oil and does not absorb water.
During centrifugation, physiological saline with the mass solubility of 0.9% and the same volume is added, the physiological saline is matched with the centrifugal pretreatment mechanism 4 to centrifugally rotate, meanwhile, the upper separation mechanism 5 and the lower separation mechanism 5 synchronously work, the rotating bearing 51 rotates to synchronously drive the lower separation structure 52 to rotate, at the moment, the high-speed separation shaft 55 and the spiral surrounding layer 56 synchronously keep rotating, an adsorption separation plate 57 is arranged on the outer surface of the spiral surrounding layer 56, the lower separation structure 52 is honeycomb-shaped, the lower separation structure 52 is made of sponge, and when the lower separation structure 52 rotates, the sponge of the lower separation structure 52 synchronously adsorbs saline in the centrifugal cavity 46; at this time, the surface of the adsorption separation plate 57 is coated with clear oil and does not absorb water, and the cells centrifuged by the centrifugal pretreatment mechanism 4 are adsorbed by the spiral arrangement of the spiral surrounding layer 56, so that the separation and collection of the centrifuged cells are realized, and the pretreatment of the stem cells is realized.
As shown in fig. 1 to 12, in one embodiment, a stem cell pretreatment method includes the steps of:
s1: stem cell crushing, namely crushing a tissue sample rich in stem cells;
s2: stem cell elutriation;
s3: the stem cells were centrifuged.
And (S1): crushing stem cells; s2: stem cell elutriation; s3: the stem cell centrifugation specifically comprises the following steps:
s1: the method for crushing the tissue sample rich in the stem cells comprises the following steps:
when in use, the stem cell tissue sample is firstly placed on the crushing blanking dish 22, the outer surface of the crushing blanking dish 22 is provided with a crushing blanking hole 23, the stem cell tissue sample leaks from the crushing blanking hole 23 and enters the inner cavity of the cylinder 21 of the crushing mechanism, a crushing blade 25 is arranged in the inner cavity, the crushing blade 25 rotates through the rotation of a crushing shaft center 24, the stem cell tissue sample is preliminarily crushed through the high-speed rotation of the crushing blade 25, and the crushed stem cell tissue sample enters the filtering and elutriating pretreatment mechanism 3;
s2: the stem cell elutriation comprises the following steps:
the crushed stem cell tissue sample immediately reaches an elutriation discharging dish 32 above a filtering and elutriation pretreatment mechanism 3, primary filtration is carried out through an elutriation discharging hole 33, the stem cell is prevented from being blocked during simultaneous discharging, the stem cell enters a barrel 31 of the elutriation mechanism and rotates through an elutriation shaft center 34, the elutriation shaft center 34 immediately drives a connecting rod 35 to rotate, the connecting rod 35 immediately drives an elutriation plate 36 to rotate, the stem cell and the tissue sample are efficiently elutriated through the rotation of the elutriation plate 36, and the stem cell obtained after elutriation flows out of an elutriation cavity 37, so that the purpose of elutriation of the stem cell is realized;
s3: the stem cell centrifugation comprises the following steps:
the stem cells after elutriation immediately enter a centrifugal blanking dish 42 above a centrifugal pretreatment mechanism 4, the stem cells are filtered through a centrifugal blanking hole 43 above the centrifugal blanking dish 42, the stem cells immediately enter a centrifugal cavity 46, the centrifugal fan 45 in a mounting base 44 is started, the centrifugal fan 45 immediately drives the stem cells in the centrifugal cavity 46 to rotate for high-speed centrifugation, meanwhile, a rotating rail 47 also keeps synchronous rotation, a vibrating block 48 connected to the inner side of the rotating rail 47 continuously rubs the inner wall of the centrifugal cavity 46, resonance occurs on the inner wall of the centrifugal cavity 46, and high-speed centrifugation of the stem cells is realized by matching with the high-speed centrifugation of the centrifugal fan 45;
during centrifugation, physiological saline with the mass solubility of 0.9% and the same volume is added, the physiological saline is matched with the centrifugal pretreatment mechanism 4 to centrifugally rotate, meanwhile, the upper separation mechanism 5 and the lower separation mechanism 5 synchronously work, the rotating bearing 51 rotates to synchronously drive the lower separation structure 52 to rotate, at the moment, the high-speed separation shaft 55 and the spiral surrounding layer 56 synchronously keep rotating, an adsorption separation plate 57 is arranged on the outer surface of the spiral surrounding layer 56, the lower separation structure 52 is honeycomb-shaped, the lower separation structure 52 is made of sponge, and when the lower separation structure 52 rotates, the sponge of the lower separation structure 52 synchronously adsorbs saline in the centrifugal cavity 46; at this time, the surface of the adsorption separation plate 57 is coated with boiled oil and does not absorb water, and the cells centrifuged by the centrifugal pretreatment mechanism 4 are adsorbed by the spiral arrangement of the spiral surrounding layer 56, so that the centrifugal cells are separated and collected, and the stem cells are pretreated.
For the convenience of understanding the above technical solutions of the present invention, the following detailed description is made of the working principle or the operation mode of the present invention in the practical process:
the working principle of the stem cell pretreatment device is as follows: when using, place stem cell tissue sample on smashing unloading ware 22 earlier, smash unloading hole 23 has been seted up on the surface of smashing unloading ware 22, the stem cell leaks down from smashing unloading hole 23, enter into the inner chamber of rubbing crusher structure barrel 21, be provided with crushing blade 25 in the inner chamber, crushing blade 25 rotates through the rotation of smashing axle center 24, high-speed rotation through crushing blade 25, carry out preliminary crushing to stem cell tissue sample, the stem cell after smashing enters into and filters in the elutriation preprocessing mechanism 3.
The crushed stem cell tissue sample immediately reaches an elutriation discharging dish 32 above a filtering and elutriation pretreatment mechanism 3, primary filtration is carried out through an elutriation discharging hole 33, the stem cell tissue sample is prevented from being blocked during simultaneous discharging, the stem cell tissue sample enters a barrel 31 of the elutriation mechanism, the elutriation shaft center 34 rotates, the elutriation shaft center 34 immediately drives a connecting rod 35 to rotate, the connecting rod 35 immediately drives an elutriation plate 36 to rotate, the stem cell tissue sample is efficiently elutriated through rotation of the elutriation plate 36, and the elutriated stem cell flows out of an elutriation cavity 37, so that the purpose of elutriating the stem cell is achieved.
The stem cells after elutriation immediately enter a centrifugal blanking dish 42 above a centrifugal pretreatment mechanism 4, the stem cells immediately enter a centrifugal cavity 46 after being filtered through a centrifugal blanking hole 43 above the centrifugal blanking dish 42, the stem cells immediately enter the centrifugal cavity 46, the centrifugal fan 45 in an installation base 44 is started, the centrifugal fan 45 immediately drives the stem cells in the centrifugal cavity 46 to rotate for high-speed centrifugation, meanwhile, a rotation rail 47 also keeps synchronous rotation, a vibration block 48 connected to the inner side of the rotation rail 47 continuously rubs the inner wall of the centrifugal cavity 46 to enable the inner wall of the centrifugal cavity 46 to resonate, and high-efficiency centrifugation of the stem cells is realized by matching with high-speed centrifugation of the centrifugal fan 45; during centrifugation, physiological saline with the mass solubility of 0.9% and the same volume is added, the physiological saline is matched with the centrifugal pretreatment mechanism 4 to rotate in a centrifugal mode, meanwhile, the upper separation mechanism 5 and the lower separation mechanism 5 work synchronously, the rotating bearing 51 rotates to synchronously drive the lower separation structure 52 to rotate, at the moment, the high-speed separation shaft 55 and the spiral surrounding layer 56 also synchronously keep rotating, an adsorption separation plate 57 is arranged on the outer surface of the spiral surrounding layer 56, the lower separation structure 52 is cellular, the lower separation structure 52 is made of sponge, and when the lower separation structure 52 rotates, the sponge of the lower separation structure 52 synchronously adsorbs saline in the centrifugal cavity 46; at this time, the surface of the adsorption separation plate 57 is coated with clear oil and does not absorb water, and the cells centrifuged by the centrifugal pretreatment mechanism 4 are adsorbed by the spiral arrangement of the spiral surrounding layer 56, so that the separation and collection of the centrifuged cells are realized, and the pretreatment of the stem cells is realized.
Finally, it should be noted that: although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still make modifications to the technical solutions described in the foregoing embodiments, or make equivalent substitutions and improvements to part of the technical features of the foregoing embodiments, and any modifications, equivalent substitutions and improvements made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (12)
1. A stem cell pretreatment device, comprising:
a device body (1);
a pulverization pretreatment mechanism (2) for pulverizing the tissue sample rich in stem cells;
a filtration and elutriation pretreatment mechanism (3) for elutriating the crushed stem cell tissue sample;
a centrifugal pretreatment mechanism (4) for centrifuging the stem cells obtained by elutriation;
and an upper and lower separation means (5) for layering the stem cells after the centrifugation.
2. The stem cell pretreatment device according to claim 1, characterized in that: the crushing pretreatment mechanism (2) comprises:
a crushing mechanism cylinder (21) which is a cylindrical cylinder;
the crushing blanking vessel (22) is concave and is arranged at the top end of the crushing mechanism cylinder body (21);
and the crushing and blanking hole (23) is formed in the outer surface of the crushing and blanking vessel (22).
3. The stem cell pretreatment device according to claim 2, characterized in that: the crushing pretreatment mechanism (2) further comprises:
a grinding shaft (24);
and a crushing blade (25) which is arranged on the outer surface of the crushing shaft center (24) in a surrounding manner, rotates along with the rotation of the crushing shaft center (24), and is arranged inside the crushing mechanism cylinder (21).
4. The stem cell pretreatment device according to claim 2, characterized in that: the filtration and elutriation pretreatment mechanism (3) comprises:
the elutriation mechanism barrel (31) is a cylindrical barrel and is connected below the crushing mechanism barrel (21);
the elutriation blanking dish (32) is concave, is arranged at the top end of the elutriation mechanism cylinder body (31) and is positioned below the crushing mechanism cylinder body (21);
and the elutriation blanking hole (33) is formed in the outer surface of the elutriation blanking dish (32), and the aperture of the elutriation blanking hole (33) is smaller than that of the crushing blanking hole (23).
5. The stem cell pretreatment device according to claim 4, characterized in that: the filtering and elutriating pretreatment mechanism (3) further comprises:
an elutriation axis (34) which is a main driving source of the filtration and elutriation pretreatment mechanism (3);
a connecting rod (35) connected to the outside of the elutriation axis (34);
the elutriation plate (36) is connected to the top end of the connecting rod (35) and is a rectangular plate;
a elutriation chamber (37), a cavity formed between adjacent elutriation plates (36).
6. The stem cell pretreatment device according to claim 1, characterized in that: the centrifugal pretreatment mechanism (4) comprises:
the centrifugal mechanism barrel (41) is a cylindrical barrel and is connected below the elutriation mechanism barrel (31);
the centrifugal blanking dish (42) is concave, is arranged at the top end of the centrifugal mechanism cylinder body (41) and is positioned below the elutriation mechanism cylinder body (31);
the centrifugal blanking hole (43) is formed in the outer surface of the centrifugal blanking dish (42), and the aperture of the centrifugal blanking hole (43) is smaller than that of the elutriation blanking hole (33).
7. The stem cell pretreatment device according to claim 6, characterized in that: the centrifugal pretreatment mechanism (4) further comprises:
a mounting base (44) mounted at the bottom end of the centrifugal mechanism cylinder (41)
A centrifugal fan (45) mounted inside the mounting base (44);
an eccentric chamber (46);
the rotating track (47) is wound on the outer surface of the centrifugal cavity (46) in a circumferential shape;
and the vibration block (48) is attached to the inner side of the rotating track (47) and is overlapped with the centrifugal cavity (46).
8. The stem cell pretreatment device according to claim 1, characterized in that: the diameter of the holes of the crushing blanking holes (23) is larger than that of the elutriation blanking holes (33), and the diameter of the holes of the elutriation blanking holes (33) is larger than that of the centrifugal blanking holes (43).
9. The stem cell pretreatment device according to claim 1, characterized in that: the upper and lower separation mechanism (5) is arranged in the centrifugal pretreatment mechanism (4) and penetrates through the inside of the centrifugal pretreatment mechanism (4) from top to bottom.
10. The stem cell pretreatment device according to claim 1, characterized in that: the upper and lower separating mechanism (5) includes:
a rotary bearing (51) arranged at the bottom end of the up-and-down separating mechanism (5);
a lower separation structure (52) connected to the rotary bearing (51);
the connecting strip (53) is connected above the lower-layer separation structure (52) and is installed in an arc shape;
and the mounting table (54) is connected with the connecting strip (53) and is arranged at the top end of the connecting strip (53).
11. The stem cell pretreatment device according to claim 10, characterized in that: the upper and lower separating mechanism (5) further comprises:
a high-speed release shaft (55);
a spiral winding layer (56) wound on the outer surface of the high-speed separation shaft (55) and mounted in a spiral winding manner;
an adsorption separator plate (57) disposed on the spiral wrap (56);
the lower-layer separation structure (52) is honeycomb-shaped, the lower-layer separation structure (52) is made of sponge, and clear oil is smeared on the surface of the adsorption separation plate (57).
12. A stem cell pretreatment method applied to the stem cell pretreatment device according to any one of claims 1 to 11, characterized in that: the processing method comprises the following steps:
s1: comminuting the tissue sample enriched for stem cells;
s2: stem cell elutriation;
s3: the stem cells were centrifuged.
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