CN115322875B - 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 the cell processing technology, and discloses a stem cell pretreatment device and a stem cell pretreatment method, wherein the stem cell pretreatment device comprises a device main body; a crush pretreatment mechanism for crushing a tissue sample rich in stem cells; filtering the elutriation pretreatment mechanism to elutriate the crushed stem cells; and the centrifugal pretreatment mechanism is used for carrying out centrifugal treatment on the stem cells. The stem cells obtained after elutriation enter a centrifugal blanking vessel above a centrifugal pretreatment mechanism immediately, are filtered through a centrifugal blanking hole above the centrifugal blanking vessel, enter a centrifugal cavity immediately, and drive stem cells in the centrifugal cavity to rotate immediately by starting a centrifugal fan in an installation base to carry out high-speed centrifugation, meanwhile, a rotating track keeps synchronous rotation, and a vibration block connected with the inner side of the rotating track does not stop rubbing 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, so that the high-efficiency centrifugation of the stem cells is realized, and the pretreatment of the stem cells is realized.
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 various mature cells, including: adipocytes, chondrocytes, osteocytes, tenocytes and myocytes. These properties, as well as their developmental plasticity, have attracted considerable attention from the scientific community as they can be used in regenerative medicine research.
The traditional umbilical cord stem cells are generally cultured by adopting a culture dish after pretreatment is carried out on human umbilical cord tissues to obtain purer human umbilical cord mesenchymal stem cells. Enrichment of stem cells has been performed in several ways. Typical methods include blocking, enzymatic hydrolysis, density gradient, panning, centrifugation, hypotonic shock to cause red blood cell lysis, and various combinations of these methods. For example, purification of stem cells from bone marrow requires removal of erythrocytes and granulocytes, which is often accomplished by density gradient centrifugation. Each of these methods has the disadvantage that one of them requires a laborious washing step after the enrichment step, typically washing the cells by repeated methods. This step typically involves placing the enriched cell suspension into a centrifuge tube and using the centrifuge to pellet the cells to the bottom of the tube. The tube was removed from the centrifuge and the supernatant was decanted from the pelleted cells. Wash solution was added to the tube and the cell pellet was resuspended. These steps are typically repeated 2 to 4 times. The use process is tedious, and is often completed through the cooperation of a plurality of groups of mechanisms, and cannot be realized through a group of devices.
We have therefore proposed stem cell pretreatment devices and methods to address these issues.
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, which solve the problems that the prior device is complicated in use process and is often completed through the cooperation of a plurality of groups of mechanisms and cannot be realized through a group of devices.
The technical scheme is as follows: in order to achieve the above purpose, the present invention provides the following technical solutions:
a stem cell pretreatment device comprising:
a device body;
a crush pretreatment mechanism for crushing a tissue sample rich in (containing) stem cells;
filtering the elutriation pretreatment mechanism to elutriate the stem cell tissue samples after being crushed;
the centrifugal pretreatment mechanism is used for carrying out centrifugal treatment on the stem cells;
and an up-down separation mechanism for layering the stem cells after the centrifugation.
Further preferable technical solution, the crushing pretreatment mechanism includes:
the crushing mechanism cylinder is a cylindrical cylinder;
the crushing blanking vessel is concave downwards and is arranged at the top end of the crushing mechanism cylinder;
and the crushing blanking hole is formed in the outer surface of the crushing blanking dish.
The crushing pretreatment mechanism further includes:
crushing the axle center;
the crushing blade is arranged on the outer surface of the crushing axis in a surrounding manner, rotates along with the rotation of the crushing axis and is arranged in the crushing mechanism cylinder.
Firstly, placing a stem cell tissue sample on a crushing blanking dish, arranging a crushing blanking hole on the outer surface of the crushing blanking dish, leaking the stem cell tissue sample from the crushing blanking hole, entering the inner cavity of a crushing mechanism barrel, arranging a crushing blade in the inner cavity, rotating the crushing blade through the rotation of a crushing shaft center, primarily crushing the stem cell tissue sample through the high-speed rotation of the crushing blade, and entering the filtering and washing pretreatment mechanism after crushing the stem cell tissue sample.
Further preferred technical solution, the filtering elutriation pretreatment mechanism includes:
the washing mechanism cylinder is a cylindrical cylinder and is connected below the crushing mechanism cylinder;
the elutriation blanking dish is concave downwards and is arranged at the top end of the elutriation mechanism cylinder and is positioned below the crushing mechanism cylinder;
and the elutriation blanking hole is formed in the outer surface of the elutriation blanking dish, and the aperture of the elutriation blanking hole is smaller than that of the crushing blanking hole.
The filter panning 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;
and a panning chamber, a cavity being formed between the adjacent panning plates.
The crushed stem cell tissue samples immediately reach the washing and discharging dish above the filtering and washing pretreatment mechanism, the preliminary filtering is carried out through the washing and discharging hole, the stem cell tissue samples are prevented from being blocked during the simultaneous discharging, the stem cell tissue samples enter the washing mechanism barrel body and rotate through the washing axle center, the washing axle center immediately drives the connecting rod to rotate, the connecting rod immediately drives the washing plate to rotate, the stem cells are efficiently washed through the rotation of the washing plate, and the stem cells obtained after washing flow out of the washing cavity, so that the aim of washing the stem cells is fulfilled.
Further preferred technical solution, the centrifugal pretreatment mechanism includes:
the centrifugal mechanism cylinder is a cylindrical cylinder and is connected below the elutriation mechanism cylinder;
a centrifugal blanking vessel which is concave and 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 dish, and the aperture of the centrifugal blanking hole is smaller than that of the elutriation blanking hole.
The centrifugal pretreatment mechanism further includes:
the mounting base is arranged at the bottom end of the centrifugal mechanism cylinder;
a centrifugal fan mounted inside the mounting base;
a centrifugal chamber;
the rotating rail is circumferentially wound on the outer surface of the centrifugal cavity;
and the vibration block is attached to the inner side of the rotating track and is overlapped with the centrifugal cavity.
The diameter of the holes of the crushing blanking holes is larger than that of the holes 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 enter a centrifugal blanking vessel above a centrifugal pretreatment mechanism immediately, are filtered through a centrifugal blanking hole above the centrifugal blanking vessel, enter a centrifugal cavity immediately, and drive stem cells in the centrifugal cavity to rotate immediately by starting a centrifugal fan in an installation base to carry out high-speed centrifugation, meanwhile, a rotating track keeps synchronous rotation, and a vibration block connected with the inner side of the rotating track does not stop rubbing the inner wall of the centrifugal cavity, so that resonance occurs on the inner wall of the centrifugal cavity, and high-speed centrifugation of the stem cells is realized by matching with the high-speed centrifugation of the centrifugal fan.
According to a further preferable technical scheme, the upper and lower separation mechanisms are arranged in the centrifugal pretreatment mechanism and penetrate through the centrifugal pretreatment mechanism up and down;
the up-down separation 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 is 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 honeycomb-shaped, and the lower layer separation structure is made of sponge materials. The surface of the adsorption separation plate is smeared with clear oil and does not absorb water.
During centrifugation, adding normal saline with the mass solubility of 0.9% with the same volume, and matching with a centrifugal pretreatment mechanism for centrifugal rotation, simultaneously, synchronously working an upper separation mechanism and a lower separation mechanism, rotating a rotating bearing, and synchronously driving a lower separation structure to rotate, wherein at the moment, a high-speed separation shaft and a spiral surrounding layer also synchronously keep rotating, an adsorption separation plate is arranged on the outer surface of the spiral surrounding layer, the lower separation structure is honeycomb-shaped, the lower separation structure is made of sponge materials, and the sponge materials of the lower separation structure synchronously adsorb the saline in a centrifugal cavity while the lower separation structure rotates; at this time, the surface of the adsorption separation plate is smeared with clear oil and does not absorb water, and cells after being centrifuged by the centrifugal pretreatment mechanism are adsorbed through the spiral arrangement of the spiral surrounding layer, so that separation and collection of centrifugal cells are realized, and pretreatment of stem cells is realized.
Further preferred embodiments provide a stem cell pretreatment method, the treatment method comprising the steps of:
s1: comminuting a tissue sample enriched in stem (containing) cells;
s2: stem cell panning;
s3: stem cells are centrifuged.
The S1: comminuting a stem cell enriched tissue sample; s2: stem cell panning; s3: stem cell centrifugation specifically comprises the following steps:
s1: comminuting the stem cell enriched tissue sample comprises the steps of:
when the filter washing pretreatment device is used, a tissue sample containing stem cells is firstly placed on a grinding and blanking vessel, a grinding and blanking hole is formed in the outer surface of the grinding and blanking vessel, the stem cell tissue sample leaks from the grinding and blanking hole and enters the inner cavity of the barrel of the grinding mechanism, a grinding blade is arranged in the inner cavity and rotates through the rotation of a grinding shaft center, the tissue sample containing the stem cells is primarily ground through the high-speed rotation of the grinding blade, and the tissue sample containing the stem cells after grinding enters the filter washing pretreatment mechanism;
s2: stem cell panning includes the steps of:
the crushed stem cell tissue samples immediately arrive at a washing and discharging dish above the filtering and washing pretreatment mechanism, preliminary filtering is carried out through a washing and discharging hole, the stem cell tissue samples are prevented from being blocked at the same time, the stem cell tissue samples enter a washing mechanism barrel body and rotate through a washing axle center, the washing axle center immediately drives a connecting rod to rotate, the connecting rod immediately drives a washing plate to rotate, the stem cell tissue samples are efficiently washed through the rotation of the washing plate, and the stem cells after washing flow out of a washing cavity, so that the aim of washing the stem cells is fulfilled;
s3: stem cell centrifugation comprises the steps of:
the method comprises the steps that the elutriated stem cells immediately enter a centrifugal blanking vessel above a centrifugal pretreatment mechanism, are filtered through a centrifugal blanking hole above the centrifugal blanking vessel, enter a centrifugal cavity immediately, and immediately drive stem cells in the centrifugal cavity to rotate for high-speed centrifugation by starting a centrifugal fan in an installation base, meanwhile, a rotating track keeps synchronous rotation, and a vibration block connected with the inner side of the rotating track does not stop rubbing 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, so that the high-efficiency centrifugation of the stem cells is realized;
during centrifugation, adding normal saline with the mass solubility of 0.9% with the same volume, and matching with a centrifugal pretreatment mechanism for centrifugal rotation, simultaneously, synchronously working an upper separation mechanism and a lower separation mechanism, rotating a rotating bearing, and synchronously driving a lower separation structure to rotate, wherein at the moment, a high-speed separation shaft and a spiral surrounding layer also synchronously keep rotating, an adsorption separation plate is arranged on the outer surface of the spiral surrounding layer, the lower separation structure is honeycomb-shaped, the lower separation structure is made of sponge materials, and the sponge materials of the lower separation structure synchronously adsorb the saline in a centrifugal cavity while the lower separation structure rotates; at this time, the surface of the adsorption separation plate is smeared with clear oil and does not absorb water, and cells after being centrifuged by the centrifugal pretreatment mechanism are adsorbed through the spiral arrangement of the spiral surrounding layer, so that separation and collection of centrifugal cells are realized, and pretreatment of stem cells is realized.
Compared with the prior art, the technical scheme of the invention has the following beneficial effects:
1. according to the stem cell pretreatment device and the stem cell pretreatment method, the stem cell tissue sample is firstly placed on the crushing blanking vessel, 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 the inner cavity of the crushing mechanism cylinder, the crushing blade is arranged in the inner cavity and rotates through the rotation of the crushing shaft center, and the stem cell tissue sample is primarily crushed through the high-speed rotation of the crushing blade, so that the aim of crushing the stem cell tissue sample is fulfilled.
2. According to the stem cell pretreatment device and method, the preliminary filtration is carried out through the elutriation blanking hole, the stem cell tissue sample is prevented from being blocked during blanking, the stem cell tissue sample enters the elutriation mechanism barrel and rotates through the elutriation axle center, the elutriation axle center immediately drives the connecting rod to rotate, the connecting rod immediately drives the elutriation plate to rotate, the stem cells are efficiently elutriated through the rotation of the elutriation plate, and the stem cells obtained after the elutriation flow out of the elutriation cavity, so that the aim of elutriating the stem cells is fulfilled.
3. According to the stem cell pretreatment device and the stem cell pretreatment method, stem cells obtained after washing enter a centrifugal blanking vessel above a centrifugal pretreatment mechanism immediately, are filtered through a centrifugal blanking hole above the centrifugal blanking vessel, enter a centrifugal cavity immediately, and are driven to rotate by a centrifugal fan in a mounting base immediately to carry out high-speed centrifugation by starting the centrifugal fan, meanwhile, a rotating track is kept to rotate synchronously, 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 stem cells is realized by matching with the high-speed centrifugation of the centrifugal fan.
4. According to the stem cell pretreatment device and the stem cell pretreatment method, physiological saline with the mass solubility of 0.9% with the equivalent volume is added during centrifugation, centrifugal pretreatment mechanism is matched for centrifugal rotation, an adsorption separation plate is arranged on the outer surface of a spiral surrounding layer, a lower separation structure is honeycomb-shaped, the lower separation structure is made of sponge materials, and the sponge materials of the lower separation structure synchronously adsorb the saline in a centrifugal cavity during rotation of the lower separation structure; at this time, the surface of the adsorption separation plate is smeared with clear oil and does not absorb water, and cells after being centrifuged by the centrifugal pretreatment mechanism are adsorbed through the spiral arrangement of the spiral surrounding layer, so that separation and collection of centrifugal cells are realized, and pretreatment of stem cells is realized.
Drawings
The accompanying drawings are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate the invention and together with the embodiments of the invention, serve to explain the invention. In the drawings:
FIG. 1 is a schematic diagram showing the overall structure of a stem cell pretreatment device according to the present invention.
FIG. 2 is a schematic diagram showing the split state of the structure in FIG. 1 of the stem cell pretreatment device according to the present invention.
FIG. 3 is a schematic perspective view of the stem cell pretreatment device of the present invention according to the embodiment of FIG. 2.
FIG. 4 is a schematic diagram showing the connection structure of the pulverizing and pretreatment mechanism of the stem cell pretreatment device of the present invention.
FIG. 5 is a schematic diagram showing the connection structure of the filtering and panning pretreatment mechanism of the stem cell pretreatment device of the present invention.
FIG. 6 is a schematic diagram showing the connection structure of the elutriation plate of the stem cell pretreatment device of the present invention.
FIG. 7 is a schematic diagram showing the connection structure of the centrifugal pretreatment mechanism of the stem cell pretreatment device of the present invention.
FIG. 8 is a schematic diagram showing the connection structure of the centrifugal chamber of the stem cell pretreatment device of the present invention.
FIG. 9 is a schematic diagram showing the structure of the stem cell pretreatment device of the present invention in a disassembled state in FIG. 8.
FIG. 10 is a schematic cross-sectional view showing the structure of the stem cell pretreatment device of the present invention in FIG. 9.
FIG. 11 is a schematic diagram showing the connection structure of the upper and lower separating mechanisms in the stem cell pretreatment device of the present invention.
FIG. 12 is an enlarged schematic view of the stem cell pretreatment device of the present invention in the region A of FIG. 11.
In the figure: 1. a device body;
2. a crushing pretreatment mechanism; 21. a crushing mechanism cylinder; 22. crushing a blanking dish; 23. crushing the blanking hole; 24. crushing the axle center; 25. crushing blade.
3. Filtering and washing pretreatment mechanism; 31. a panning mechanism cylinder; 32. washing the blanking dish; 33. washing the blanking hole; 34. washing the axle center; 35. a connecting rod; 36. a elutriation plate; 37. the cavity was panned.
4. A centrifugal pretreatment mechanism; 41. a centrifugal mechanism cylinder; 42. centrifuging the blanking vessel; 43. centrifuging the blanking hole; 44. a mounting base; 45. a centrifugal fan; 46. a centrifugal chamber; 47. rotating the track; 48. and (5) vibrating the block.
5. An up-down separating mechanism; 51. a rotating bearing; 52. a lower layer separation structure; 53. a connecting strip; 54. a mounting table; 55. a high-speed separation shaft; 56. a spiral surrounding layer; 57. adsorption separation plate.
Description of the embodiments
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
As shown in fig. 1 to 3, in one embodiment, the stem cell pretreatment device includes:
a device main body 1;
a pulverizing pretreatment mechanism 2 for pulverizing stem cells;
a filtering and panning pretreatment mechanism 3 for panning the crushed stem cells;
a centrifugal pretreatment mechanism 4 for centrifuging stem cells;
and an up-down separation mechanism 5 for separating the stem cells after centrifugation.
As shown in fig. 4, in one embodiment, the stem cell pretreatment device includes:
a device main body 1;
a pulverizing pretreatment mechanism 2 for pulverizing stem cells;
a filtering and panning pretreatment mechanism 3 for panning the crushed stem cells;
a centrifugal pretreatment mechanism 4 for centrifuging stem cells;
an up-down separation mechanism 5 for layering stem cells after centrifugation;
the pulverization pretreatment mechanism 2 comprises:
the crushing mechanism cylinder 21 is a cylindrical cylinder;
a pulverizing and discharging dish 22 having a concave shape and mounted on the top end of the pulverizing mechanism cylinder 21;
a crushing blanking hole 23 is formed on the outer surface of the crushing blanking dish 22.
The pulverization pretreatment mechanism 2 further comprises:
a crushing shaft center 24;
the pulverizing blade 25 is circumferentially provided on the outer surface of the pulverizing shaft 24, rotates with the rotation of the pulverizing shaft 24, and is provided inside the pulverizing mechanism cylinder 21.
The stem cells are firstly placed on a crushing blanking dish 22, crushing blanking holes 23 are formed in the outer surface of the crushing blanking dish 22, the stem cells leak from the crushing blanking holes 23 and enter the inner cavity of the crushing mechanism barrel 21, crushing blades 25 are arranged in the inner cavity, the crushing blades 25 rotate through the rotation of a crushing shaft center 24, the stem cells are primarily crushed through the high-speed rotation of the crushing blades 25, and the crushed stem cells enter the filtering elutriation pretreatment mechanism 3.
As shown in fig. 5 to 6, in one embodiment, the stem cell pretreatment device includes:
a device main body 1;
a crush pretreatment mechanism 2 for crushing a tissue sample rich in stem cells;
a filtering and panning pretreatment mechanism 3 for panning the crushed stem cell tissue sample;
a centrifugal pretreatment mechanism 4 for centrifuging stem cells;
an up-down separation mechanism 5 for layering stem cells after centrifugation;
the filtering elutriation pretreatment mechanism 3 comprises:
the washing mechanism cylinder 31 is a cylindrical cylinder and is connected below the crushing mechanism cylinder 21;
a washing and discharging dish 32 having a concave shape and mounted on the top end of the washing mechanism cylinder 31 below the pulverizing mechanism cylinder 21;
and a elutriation blanking hole 33 which is formed on the outer surface of the elutriation blanking plate 32, wherein the aperture of the elutriation blanking hole 33 is smaller than that of the crushing blanking hole 23.
The filtering elutriation pretreatment mechanism 3 further comprises:
the elutriation axis 34 is a main driving source of the filtering elutriation pretreatment mechanism 3;
a connecting rod 35 connected to the outside of the washing shaft 34;
the washing plate 36 is connected to the top end of the connecting rod 35 and is a rectangular plate;
a panning chamber 37, a cavity formed between the adjacent panning plates 36.
The crushed stem cell tissue samples then reach the washing and blanking vessel 32 above the filtering and washing pretreatment mechanism 3, the preliminary filtering is carried out through the washing and blanking holes 33, the stem cells are prevented from being blocked at the same time, the stem cells enter the washing mechanism barrel 31, the washing axis 34 rotates, the washing axis 34 drives the connecting rod 35 to rotate, the connecting rod 35 drives the washing plate 36 to rotate, the stem cells are efficiently washed through the rotation of the washing plate 36, and the washed stem cells flow out of the washing cavity 37, so that the aim of washing the stem cells is fulfilled.
As shown in fig. 7 to 10, in one embodiment, the stem cell pretreatment device includes:
a device main body 1;
a crush pretreatment mechanism 2 for crushing a stem cell tissue sample;
a filtering and panning pretreatment mechanism 3 for panning the crushed stem cells;
a centrifugal pretreatment mechanism 4 for centrifuging stem cells;
the centrifugal pretreatment mechanism 4 includes:
an up-down separation mechanism 5 for layering stem cells after centrifugation;
a centrifugal mechanism cylinder 41 having a cylindrical shape and connected to the lower side of the elutriation mechanism cylinder 31;
a centrifugal blanking plate 42 having a concave shape and mounted on the top end of the centrifugal mechanism cylinder 41 below the elutriation mechanism cylinder 31;
and a centrifugal blanking hole 43 formed on the outer surface of the centrifugal blanking plate 42, wherein the aperture of the centrifugal blanking hole 43 is smaller than that of the elutriation blanking hole 33.
The centrifugal pretreatment mechanism 4 further includes:
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;
a centrifugal chamber 46;
a rotation rail 47 circumferentially wound around the outer surface of the centrifugal chamber 46;
and a vibration block 48 attached to the inner side of the rotation rail 47 and overlapping the centrifugal chamber 46.
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.
The elutriated stem cells immediately enter the centrifugal blanking dish 42 above the centrifugal pretreatment mechanism 4, the stem cells are filtered through the centrifugal blanking hole 43 above the centrifugal blanking dish 42, then enter the centrifugal cavity 46, the centrifugal fan 45 in the 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, the rotating track 47 also keeps synchronous rotation, the vibration block 48 connected to the inner side of the rotating track 47 does not stop rubbing against 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 centrifugal fan 45 is matched, so that high-efficiency centrifugation of the stem cells is realized.
As shown in fig. 11 to 12, in one embodiment, the upper and lower separation mechanism 5 is provided inside the centrifugal pretreatment mechanism 4, penetrating up and down inside the centrifugal pretreatment mechanism 4;
the vertical separation mechanism 5 includes:
a rotation bearing 51 provided at the bottom end of the vertical separation mechanism 5;
a lower layer separation structure 52 connected to the rotation bearing 51;
the connecting strip 53 is connected above the lower layer separation structure 52 and is installed in an arc shape;
a mounting table 54 connected to the connection bar 53 and provided at the top end of the connection bar 53;
a high-speed separation shaft 55;
a spiral surrounding layer 56 which is wound on the outer surface of the high-speed separation shaft 55 and is spirally wound;
an adsorption partition plate 57 is provided on the spiral surrounding layer 56.
The lower layer separation structure 52 is honeycomb-shaped, and the lower layer separation structure 52 is made of sponge material. The surface of the adsorption separation plate 57 is smeared with clear oil and does not absorb water.
During centrifugation, physiological saline with the mass solubility of 0.9% with the equivalent volume is added, and the centrifugal pretreatment mechanism 4 is matched for centrifugal rotation, meanwhile, the upper separation mechanism 5 and the lower separation mechanism 5 synchronously work, the rotating bearing 51 rotates and synchronously drives 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, the outer surface of the spiral surrounding layer 56 is provided with an adsorption separation plate 57, the lower separation structure 52 is honeycomb-shaped, the lower separation structure 52 is made of sponge materials, and the sponge materials of the lower separation structure 52 synchronously adsorb the saline in the centrifugal cavity 46 when the lower separation structure 52 rotates; at this time, the surface of the adsorption separation plate 57 is smeared with clear oil, and is not water-absorbing, and the cells after centrifugation by the centrifugal pretreatment mechanism 4 are adsorbed by the spiral arrangement of the spiral surrounding layer 56, so that separation and collection of centrifugal cells are realized, and pretreatment of 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 panning;
s3: stem cells are centrifuged.
The S1: crushing stem cells; s2: stem cell panning; s3: stem cell centrifugation specifically comprises the following steps:
s1: comminuting the stem cell enriched tissue sample comprises the steps of:
when the filter washing pretreatment device is used, a stem cell tissue sample is firstly placed on a grinding and blanking vessel 22, a grinding and blanking hole 23 is formed in the outer surface of the grinding and blanking vessel 22, the stem cell tissue sample leaks from the grinding and blanking hole 23 and enters the inner cavity of a grinding mechanism cylinder 21, a grinding blade 25 is arranged in the inner cavity, the grinding blade 25 rotates through the rotation of a grinding shaft center 24, the stem cell tissue sample is primarily ground through the high-speed rotation of the grinding blade 25, and the ground stem cell tissue sample enters the filter washing pretreatment mechanism 3;
s2: stem cell panning includes the steps of:
the crushed stem cell tissue samples immediately reach the elutriation blanking plate 32 above the filtering and elutriation pretreatment mechanism 3, the preliminary filtering is carried out through the elutriation blanking holes 33, the stem cells are prevented from being blocked at the same time, the stem cells enter the elutriation mechanism cylinder 31, the elutriation axis 34 rotates, the elutriation axis 34 immediately drives the connecting rod 35 to rotate, the connecting rod 35 immediately drives the elutriation plate 36 to rotate, the stem cells and the tissue samples are efficiently elutriated through the rotation of the elutriation plate 36, and the stem cells obtained after the elutriation flow out of the elutriation cavity 37, so that the aim of elutriating the stem cells is fulfilled;
s3: stem cell centrifugation comprises the steps of:
the elutriated stem cells immediately enter a centrifugal blanking vessel 42 above a centrifugal pretreatment mechanism 4, are filtered through a centrifugal blanking hole 43 above the centrifugal blanking vessel 42, enter a centrifugal cavity 46 immediately, and drive the stem cells in the centrifugal cavity 46 to rotate by starting a centrifugal fan 45 in a mounting base 44 immediately to carry out high-speed centrifugation, meanwhile, a rotating track 47 also keeps synchronous rotation, a vibration block 48 connected with the inner side of the rotating track 47 does not stop rubbing the inner wall of the centrifugal cavity 46, so that resonance occurs on the inner wall of the centrifugal cavity 46, and the high-speed centrifugation of the centrifugal fan 45 is matched, so that the high-efficiency centrifugation of the stem cells is realized;
during centrifugation, physiological saline with the mass solubility of 0.9% with the equivalent volume is added, and the centrifugal pretreatment mechanism 4 is matched for centrifugal rotation, meanwhile, the upper separation mechanism 5 and the lower separation mechanism 5 synchronously work, the rotating bearing 51 rotates and synchronously drives 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, the outer surface of the spiral surrounding layer 56 is provided with an adsorption separation plate 57, the lower separation structure 52 is honeycomb-shaped, the lower separation structure 52 is made of sponge materials, and the sponge materials of the lower separation structure 52 synchronously adsorb the saline in the centrifugal cavity 46 when the lower separation structure 52 rotates; at this time, the surface of the adsorption separation plate 57 is smeared with clear oil, and is not water-absorbing, and the cells after centrifugation by the centrifugal pretreatment mechanism 4 are adsorbed by the spiral arrangement of the spiral surrounding layer 56, so that separation and collection of centrifugal cells are realized, and pretreatment of stem cells is realized.
In order to facilitate understanding of the above technical solutions of the present invention, the following details are about the working principle or operation mode of the present invention in the actual process:
stem cell pretreatment device theory of operation: when the stem cell tissue sample is used, the stem cell tissue sample is firstly placed on the crushing blanking vessel 22, the crushing blanking hole 23 is formed in the outer surface of the crushing blanking vessel 22, stem cells leak from the crushing blanking hole 23 and enter the inner cavity of the crushing mechanism barrel 21, the crushing blade 25 is arranged in the inner cavity, the crushing blade 25 rotates through the rotation of the crushing shaft center 24, the stem cell tissue sample is primarily crushed through the high-speed rotation of the crushing blade 25, and the crushed stem cells enter the filtering and washing pretreatment mechanism 3.
The crushed stem cell tissue sample then reaches the washing and blanking vessel 32 above the filtering and washing pretreatment mechanism 3, is preliminarily filtered through the washing and blanking hole 33, and is prevented from being blocked at the same time, the stem cell tissue sample enters the washing mechanism barrel 31, the washing axis 34 rotates, the washing axis 34 drives the connecting rod 35 to rotate, the connecting rod 35 drives the washing plate 36 to rotate, the stem cell tissue sample is efficiently washed through the rotation of the washing plate 36, and the stem cells after washing flow out of the washing cavity 37, so that the aim of washing the stem cells is fulfilled.
The elutriated stem cells immediately enter a centrifugal blanking vessel 42 above a centrifugal pretreatment mechanism 4, are filtered through a centrifugal blanking hole 43 above the centrifugal blanking vessel 42, enter a centrifugal cavity 46 immediately, and drive the stem cells in the centrifugal cavity 46 to rotate by starting a centrifugal fan 45 in a mounting base 44 immediately to carry out high-speed centrifugation, meanwhile, a rotating track 47 also keeps synchronous rotation, a vibration block 48 connected with the inner side of the rotating track 47 does not stop rubbing the inner wall of the centrifugal cavity 46, so that resonance occurs on the inner wall of the centrifugal cavity 46, and the high-speed centrifugation of the centrifugal fan 45 is matched, so that the high-efficiency centrifugation of the stem cells is realized; during centrifugation, physiological saline with the mass solubility of 0.9% with the equivalent volume is added, and the centrifugal pretreatment mechanism 4 is matched for centrifugal rotation, meanwhile, the upper separation mechanism 5 and the lower separation mechanism 5 synchronously work, the rotating bearing 51 rotates and synchronously drives 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, the outer surface of the spiral surrounding layer 56 is provided with an adsorption separation plate 57, the lower separation structure 52 is honeycomb-shaped, the lower separation structure 52 is made of sponge materials, and the sponge materials of the lower separation structure 52 synchronously adsorb the saline in the centrifugal cavity 46 when the lower separation structure 52 rotates; at this time, the surface of the adsorption separation plate 57 is smeared with clear oil, and is not water-absorbing, and the cells after centrifugation by the centrifugal pretreatment mechanism 4 are adsorbed by the spiral arrangement of the spiral surrounding layer 56, so that separation and collection of centrifugal cells are realized, and pretreatment of stem cells is realized.
Finally, it should be noted that: the foregoing description is only illustrative of the preferred embodiments of the present invention, and although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that modifications may be made to the embodiments described, or equivalents may be substituted for elements thereof, and any modifications, equivalents, improvements or changes may be made without departing from the spirit and principles of the present invention.
Claims (8)
1. Stem cell preprocessing device, characterized by includes:
a device body (1);
a crush pretreatment mechanism (2) for crushing a tissue sample rich in stem cells;
a filtering and panning pretreatment mechanism (3) for panning the crushed stem cell tissue sample;
a centrifugal pretreatment mechanism (4) for centrifuging the stem cells obtained by the elutriation;
an up-down separation mechanism (5) for layering stem cells after centrifugation;
the centrifugal pretreatment mechanism (4) comprises:
a centrifugal chamber (46);
a rotating rail (47) which is wound on the outer surface of the centrifugal cavity (46) in a circumferential manner;
a vibration block (48) attached to the inner side of the rotation rail (47) and overlapping the centrifugal cavity (46);
the upper and lower separation mechanisms (5) are arranged in the centrifugal pretreatment mechanism (4) and penetrate through the centrifugal pretreatment mechanism (4) up and down;
the vertical separation mechanism (5) comprises:
a rotating bearing (51) arranged at the bottom end of the upper and lower separating mechanism (5);
a lower layer separation structure (52) connected to the rotation bearing (51);
the connecting strip (53) is connected above the lower separation structure (52) and is installed in an arc shape;
the mounting table (54) is connected with the connecting strip (53) and is arranged at the top end of the connecting strip (53);
the vertical separation mechanism (5) further comprises:
a high-speed release shaft (55);
a spiral surrounding layer (56) which is wound on the outer surface of the high-speed separation shaft (55) and is spirally wound and installed;
an adsorption separation plate (57) provided on the spiral surrounding layer (56);
the lower separation structure (52) is honeycomb, the lower separation structure (52) is made of sponge materials, and clear oil is smeared on the surface of the adsorption separation plate (57).
2. The stem cell pretreatment device according to claim 1, wherein: the pulverization pretreatment 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 the crushing blanking hole (23) is formed in the outer surface of the crushing blanking dish (22).
3. The stem cell pretreatment device according to claim 2, wherein: the pulverization pretreatment mechanism (2) further comprises:
a grinding shaft center (24);
the crushing blade (25) is circumferentially arranged on the outer surface of the crushing shaft center (24), rotates along with the rotation of the crushing shaft center (24), and is arranged in the crushing mechanism cylinder (21).
4. The stem cell pretreatment device according to claim 2, wherein: the filtering elutriation pretreatment mechanism (3) comprises:
a cylindrical washing mechanism cylinder (31) which is a cylindrical cylinder and is connected below the crushing mechanism cylinder (21);
a washing and discharging dish (32) which is concave and is arranged at the top end of the washing mechanism cylinder (31) and is positioned below the crushing mechanism cylinder (21);
and a elutriation blanking hole (33) which is formed on the outer surface of the elutriation blanking plate (32), wherein 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, wherein: the filtering elutriation pretreatment mechanism (3) further comprises:
a panning axis (34) which is a main driving source of the filtering panning pretreatment mechanism (3);
a connecting rod (35) connected to the outside of the washing shaft center (34);
the elutriation plate (36) is connected to the top end of the connecting rod (35) and is a rectangular plate;
and a washing chamber (37) formed adjacent to the cavity between the washing plates (36).
6. The stem cell pretreatment device according to claim 4, wherein: the centrifugal pretreatment mechanism (4) comprises:
a centrifugal mechanism cylinder (41) which is a cylindrical cylinder and is connected below the elutriation mechanism cylinder (31);
a centrifugal blanking vessel (42) which is concave downward and is arranged at the top end of the centrifugal mechanism cylinder (41) and is positioned below the elutriation mechanism cylinder (31);
a centrifugal blanking hole (43) which is arranged on the outer surface of the centrifugal blanking dish (42), wherein the aperture of the centrifugal blanking hole (43) is smaller than that of the elutriation blanking hole (33);
a mounting base (44) mounted at the bottom end of the centrifugal mechanism cylinder (41);
and a centrifugal fan (45) mounted inside the mounting base (44).
7. The stem cell pretreatment device according to claim 6, wherein: 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).
8. A stem cell pretreatment method applied to the stem cell pretreatment device according to any one of claims 1 to 7, characterized in that: the processing method comprises the following steps:
s1: comminuting a stem cell enriched tissue sample;
s2: stem cell panning;
s3: centrifuging stem cells;
during centrifugation, adding normal saline with the mass solubility of 0.9% with the same volume, and matching with a centrifugal pretreatment mechanism for centrifugal rotation, simultaneously, synchronously working an upper separation mechanism and a lower separation mechanism, rotating a rotating bearing, and synchronously driving a lower separation structure to rotate, wherein at the moment, a high-speed separation shaft and a spiral surrounding layer also synchronously keep rotating, an adsorption separation plate is arranged on the outer surface of the spiral surrounding layer, the lower separation structure is honeycomb-shaped, the lower separation structure is made of sponge materials, and the sponge materials of the lower separation structure synchronously adsorb the saline in a centrifugal cavity while the lower separation structure rotates; at this time, the surface of the adsorption separation plate is smeared with clear oil and does not absorb water, and cells after being centrifuged by the centrifugal pretreatment mechanism are adsorbed through the spiral arrangement of the spiral surrounding layer, so that separation and collection of centrifugal cells are realized, and pretreatment of stem cells is realized.
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