CN114540289A - Freeze-drying process for mesenchymal stem cell culture solution - Google Patents
Freeze-drying process for mesenchymal stem cell culture solution Download PDFInfo
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- CN114540289A CN114540289A CN202210136504.0A CN202210136504A CN114540289A CN 114540289 A CN114540289 A CN 114540289A CN 202210136504 A CN202210136504 A CN 202210136504A CN 114540289 A CN114540289 A CN 114540289A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0662—Stem cells
- C12N5/0668—Mesenchymal stem cells from other natural sources
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0205—Chemical aspects
- A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
- A01N1/0221—Freeze-process protecting agents, i.e. substances protecting cells from effects of the physical process, e.g. cryoprotectants, osmolarity regulators like oncotic agents
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A01N1/02—Preservation of living parts
- A01N1/0236—Mechanical aspects
- A01N1/0242—Apparatuses, i.e. devices used in the process of preservation of living parts, such as pumps, refrigeration devices or any other devices featuring moving parts and/or temperature controlling components
- A01N1/0252—Temperature controlling refrigerating apparatus, i.e. devices used to actively control the temperature of a designated internal volume, e.g. refrigerators, freeze-drying apparatus or liquid nitrogen baths
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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Abstract
The invention provides a mesenchymal stem cell culture solution freeze-drying process which is characterized by comprising the following steps: s1: screening and selecting umbilical cord donors to carry out pathological detection; taking healthy umbilical cord stem cells to perform umbilical cord mesenchymal stem cell culture, and collecting mesenchymal stem cell conditioned medium; s2: centrifuging the mesenchymal stem cell conditioned culture solution, filtering the upper suspension, adding a freeze-drying protective agent and glycerol, and uniformly stirring by ultrasonic waves; s3: putting the obtained liquid into a liquid adding port of a mesenchymal stem cell culture solution freeze-drying device; s4: waiting for the organism to be subjected to freeze-drying processing to obtain the freeze-dried powder. The invention has simple process operation steps and high production capacity, and can be used for beautifying and protecting skin; the device is simple to operate, low in cost and suitable for large-scale production, and can meet the application of the mesenchymal stem cells in the field of beauty at present.
Description
Technical Field
The invention relates to the technical field of cells, in particular to a mesenchymal stem cell culture solution freeze-drying process.
Background
Mesenchymal Stem Cells (MSCs) are pluripotent stem cells that have all of the commonality of stem cells, namely self-renewal and multipotentiality. Various growth factors that promote skin cell regeneration, including basic fibroblast growth factor (bFGF), human Epidermal Growth Factor (EGF), and Vascular Endothelial Growth Factor (VEGF), are secreted in culture. MSCs also secrete large amounts of extracellular matrix, which can increase skin health and appearance. The culture solution of MSC is collected, freeze-dried and easy to store, and can be used with skin care products to achieve the effects of beautifying and protecting skin.
However, the application of the autologous mesenchymal stem cells gradually exposes inconvenience: for example, the amplification capacity varies widely among individuals; potential risk of tumor cell contamination; the culture needs a certain time, and can not adapt to the needs of the state of illness in time, etc. These restrict the use of autologous mesenchymal stem cells. The mesenchymal stem cells bring new hopes for future regenerative medicine, and the intensive research and clinical application of the mesenchymal stem cells will certainly benefit mankind in the near future.
Transplantation of human Mesenchymal Stem Cells (MSCs) has been clinically applied in various fields including the treatment of degenerative neuropathy, neonatal cerebral ischemia and adult cerebral stroke, and the treatment of cancer, and also applied to the correction of skin defects, but the use of culture supernatant thereof in beauty and skin care has just begun.
However, most of the existing methods for producing the human mesenchymal stem cell culture solution supernatant are very troublesome, have small production capacity, small amount of various growth factors in the supernatant and poor quality, and thus the human mesenchymal stem cell culture solution supernatant produced by the existing methods cannot completely meet the existing use.
Disclosure of Invention
The invention aims to provide a mesenchymal stem cell culture solution freeze-drying process to solve the problems in the background technology.
In order to achieve the purpose, the invention adopts the following technical scheme:
a mesenchymal stem cell culture solution freeze-drying process is characterized by comprising the following steps:
s1: screening and selecting umbilical cord donors to perform pathological detection; taking healthy umbilical cord stem cells to perform umbilical cord mesenchymal stem cell culture, and collecting mesenchymal stem cell conditioned medium;
s2: centrifuging the mesenchymal stem cell conditioned medium, filtering the suspension, adding a freeze-drying protective agent and glycerol, and ultrasonically stirring uniformly;
s3: putting the obtained liquid into a liquid adding port of a mesenchymal stem cell culture solution freeze-drying device;
s4: waiting for the organism to be subjected to freeze-drying processing to obtain the freeze-dried powder.
A mesenchymal stem cell culture solution freeze-drying device is characterized by comprising a solution containing module 1, a refrigerating module 2, a powder pressing module 3, a split charging module 4, a standby module 5, a power supply module 6, a control module 7 and a machine shell 8; put liquid module 1 set up in refrigeration module 2 top, powder pressing module 3 is located partial shipment module 4 one side is used for removing extremely partial shipment module 4 top, powder pressing module 3 below is equipped with spare module 5, refrigeration module 2 one side is equipped with power module 6, refrigeration module 2 with power module 6 below is equipped with control module 7.
The liquid placing module 1 comprises a liquid placing box 11, a liquid placing box fixing clamp 12, a liquid placing box sliding block 13, a first driving motor 14, a liquid placing box moving sliding rod 15 and a liquid placing box moving track 16; the liquid placing box comprises a liquid placing box 11 and a liquid placing box fixing block 115, wherein the liquid placing box 11 comprises a first half liquid placing box 111 and a second half liquid placing box 112, a half body bulge 113 is arranged at the open end of the first half liquid placing box 111, a half body depression 114 is arranged at the open end of the second half liquid placing box 112, the half body bulge 113 is matched with the half body depression 114 in shape, the liquid placing box fixing clamp 12 clamps and fixes the liquid placing box fixing block 115, the liquid placing box sliding block 13 is welded at the bottom of the liquid placing box fixing clamp 12, the liquid placing box sliding block 13 is sleeved on a liquid placing box moving sliding rod 15, the liquid placing box moving sliding rod 15 is covered with threads, the liquid placing box moving sliding rod 15 is arranged in the liquid placing box moving rail 16, and the liquid placing box moving rail 16 is fixed above the inner part of the shell 8 through a fixing rod 9.
The refrigerating module 2 comprises a refrigerator 21, a freezing pad 22 and a freezing pad moving track 23, the refrigerator 21 is connected with the freezing pad 22, and the freezing pad 22 moves left and right through the freezing pad moving track 23; the refrigerator 21 is a semiconductor refrigerator and is provided with a refrigerator water inlet 211 and a refrigerator water outlet 212, the freezing pad 22 is provided with a first freezing connector 221 and a second freezing connector 222, the refrigerator water inlet 211 is connected with the first freezing connector 221, the refrigerator water outlet 212 is connected with the second freezing connector 222, one side of the freezing pad 22 is connected with a freezing pad sliding block 225 through a freezing pad fixing clamp 223, the freezing pad sliding block 225 is provided with a second driving motor 226, the freezing pad sliding block 225 is sleeved on a freezing pad moving sliding rod 224, and the freezing pad moving sliding rod 224 is arranged in the freezing pad moving track 23.
The powder pressing module 3 comprises a grinding pressing block 31, a stabilizing block 32, a supporting rod 33, a telescopic rod 34, a grinding pressing moving slide block 35 and a grinding pressing moving track 36, the grinding block 31 is connected with the stabilizing block 32 through a connecting block 321, the stabilizing block 32 is conical and has a stabilizing effect, one end of the supporting rod 33 is welded on the stabilizing block 32, the other end of the supporting rod 33 is connected with the telescopic rod 34, the telescopic rod 34 is a reciprocating electric push rod with the model of MNTL, the telescopic rod 34 is connected with the grinding pressure moving slide block 35, the grinding pressure moving track 36 is provided with a third driving motor 351, the grinding pressure moving slide block 35 slides on the grinding pressure moving track 36, the grinding pressure moving track 36 is a linear guide rail with the model number of TRH20B, the grinding pressure moving track 36 is fixed below the inner part of the machine shell 8 through a fixing rod 9, and the standby module 5 is arranged below the grinding pressure moving track 36.
The partial shipment module 4 includes hourglass powder board 41, drawer main part 42, partial shipment heat preservation box 43, brush cleaner 44, brush-sweeping support rod 45, brush cleaner slider 46 and brush slide bar 47, brush cleaner slider 46 set up in on the brush slide bar 47, brush cleaner slider 47 passes through support rod 45 is fixed in 8 inside below of casing, brush cleaner slider 46 with brush cleaner slider 47 is equipped with two sets ofly, is equipped with between the brush cleaner 44, brush cleaner 44 follows brush cleaner slider 46's removal and move, four position within the support rod 45 is equipped with hourglass powder board 41, hourglass powder board 41 set up in on the drawer main part 42, hourglass powder board 41 with be equipped with between the drawer main part 42 partial shipment heat preservation box 43, partial shipment heat preservation box 43 is equipped with a plurality of spaced-apart freeze-drying powder for the splendid attire freeze-drying powder.
The power module 6 is arranged on one side of the refrigerator 21, the control module 7 is arranged below the refrigerator 21 and the power module 6, the power module 6 is provided with an external port, and the control module 7 is provided with an external port; the model of the power supply module 6 is X-HX0108A, and the model of the control module 7 is CPU224 XP.
The top of the machine shell 8 is provided with a liquid filling port 82, the liquid filling port 82 is provided with a liquid filling cover 81, the top of the liquid filling cover 81 is provided with a handle 811, the bottom of the liquid filling cover is provided with two locking cover protrusions 812 with opposite positions, the liquid filling port 82 is in an inverted cone shape and is provided with an upper through hole and a lower through hole, a circle above the liquid filling port 82 is provided with a locking cover groove 822, two locking cover notches 821 are arranged above the locking cover groove 822, the front surface of the machine shell 8 is provided with an operation platform 83 and a visible window 84, the back surface of the machine shell is provided with an outer machine shell interface 86, one end side surface of the machine shell is provided with a heat dissipation hole 85, and the other end side surface of the machine shell is provided with a maintenance small window 87.
The first, second and third driving motors 14, 226 and 351 are small round motors of type 1230.
Compared with the prior art, the invention has the beneficial effects that:
the invention has simple process operation steps and high production capacity, and can be used for beautifying and protecting skin; the device is simple to operate, low in cost and suitable for large-scale production, and can meet the application of the mesenchymal stem cells in the field of beauty at present.
Drawings
FIG. 1 is a schematic view of the overall structure of the interior of a mesenchymal stem cell culture solution freeze-drying device according to the present invention;
FIG. 2 is a schematic structural diagram of a liquid-containing module of the mesenchymal stem cell culture liquid freeze-drying device according to the present invention;
FIG. 3 is a schematic diagram of the separation state of the liquid placing module of the mesenchymal stem cell culture liquid freeze-drying device according to the present invention;
FIG. 4 is a schematic structural diagram of a refrigeration module of the mesenchymal stem cell culture solution freeze-drying device according to the present invention;
FIG. 5 is a schematic structural diagram of a powder pressing module and a split charging module of the mesenchymal stem cell culture solution freeze-drying device according to the present invention;
FIG. 6 is a schematic structural diagram of a powder pressing module of the mesenchymal stem cell culture solution freeze-drying device according to the present invention;
FIG. 7 is a schematic structural diagram of a split charging module of the mesenchymal stem cell culture solution freeze-drying device according to the present invention;
FIG. 8 is a schematic structural diagram of a power module and a control module of the mesenchymal stem cell culture solution freeze-drying device according to the present invention;
fig. 9 is a schematic structural diagram of a housing of the mesenchymal stem cell culture solution freeze-drying device according to the present invention;
FIG. 10 is a second schematic structural diagram of a housing of the device for lyophilizing a mesenchymal stem cell culture solution according to the present invention;
FIG. 11 is a schematic structural view of a liquid filling port and a liquid filling cover of the mesenchymal stem cell culture solution freeze-drying device according to the present invention;
in the figure: 1. a liquid containing module; 11. a liquid placing box; 111. a first half liquid containing box; 112. a second half liquid containing box; 113. half body bulges; 114. the half bodies are sunken; 115. a liquid placing box fixing block; 12. a liquid placing box fixing clamp; 13. a liquid box slide block; 14. a first drive motor; 15. the liquid placing box moves the sliding rod; 16. a liquid containing box moving track; 2. a refrigeration module; 21. a refrigerator; 211. a water inlet of the refrigerator; 212. a water outlet of the refrigerator; 22. a freezing pad; 221. a first refrigeration interface; 222. a second refrigeration interface; 223. a freezing pad fixing clip; 224. the freezing pad moves the sliding rod; 225. a freezing pad sliding block; 226. a second drive motor; 23. a freezing pad moving track; 3. a powder pressing module; 31. grinding and pressing the block; 32. a stabilizing block; 321. connecting blocks; 33. a support bar; 34. a telescopic rod; 35. grinding and pressing the movable sliding block; 351. a third drive motor; 36. grinding and pressing the moving track; 4. a split charging module; 41. a powder leaking plate; 411. a powder leakage hole; 42. a drawer main body; 43. subpackaging the heat preservation boxes; 44. cleaning with a brush; 45. a sweeping support rod; 46. a sweeping slide block; 461. a fourth drive motor; 47. a sweeper slide bar; 5. a standby module; 6. a power supply module; 7. a control module; 8. a housing; 81. a liquid feeding cover; 811. a handle; 812. a lock cover protrusion; 82. a liquid filling port; 821. a locking cover notch; 822. a locking cover groove; 83. an operation table; 84. a visual window; 85. heat dissipation holes; 86. an outer interface of the casing; 87. maintaining the small window; 9. and (5) fixing the rod.
Detailed Description
The technical solutions of the present invention will be described clearly and completely with reference to the accompanying drawings, and it is to be understood that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments, and all other embodiments obtained by those skilled in the art without any inventive work based on the embodiments of the present invention belong to the scope of the present invention.
Examples
A mesenchymal stem cell culture solution freeze-drying process is characterized by comprising the following steps:
s1: screening and selecting umbilical cord donors to carry out pathological detection; taking healthy umbilical cord stem cells to perform umbilical cord mesenchymal stem cell culture, and collecting mesenchymal stem cell conditioned medium;
s2: centrifuging the mesenchymal stem cell conditioned culture solution, filtering the upper suspension, adding a freeze-drying protective agent and glycerol, and uniformly stirring by ultrasonic waves;
s3: putting the obtained liquid into a liquid adding port of a mesenchymal stem cell culture solution freeze-drying device;
s4: waiting for the organism to be subjected to freeze-drying processing to obtain the freeze-dried powder.
As shown in fig. 1, a mesenchymal stem cell culture solution freeze-drying device is characterized by comprising a solution storage module 1, a refrigeration module 2, a powder pressing module 3, a split charging module 4, a standby module 5, a power supply module 6, a control module 7 and a casing 8; put liquid module 1 set up in refrigeration module 2 top, powder pressing module 3 is located partial shipment module 4 one side is used for removing extremely partial shipment module 4 top, powder pressing module 3 below is equipped with spare module 5, refrigeration module 2 one side is equipped with power module 6, refrigeration module 2 with power module 6 below is equipped with control module 7.
As shown in fig. 2-3, the liquid placing module 1 includes a liquid placing box 11, a liquid placing box fixing clamp 12, a liquid placing box sliding block 13, a first driving motor 14, a liquid placing box moving slide rod 15, and a liquid placing box moving track 16; the liquid placing box comprises a liquid placing box 11 and a liquid placing box fixing block 115, wherein the liquid placing box 11 comprises a first half liquid placing box 111 and a second half liquid placing box 112, a half body bulge 113 is arranged at the open end of the first half liquid placing box 111, a half body depression 114 is arranged at the open end of the second half liquid placing box 112, the half body bulge 113 is matched with the half body depression 114 in shape, the liquid placing box fixing clamp 12 clamps and fixes the liquid placing box fixing block 115, the liquid placing box sliding block 13 is welded at the bottom of the liquid placing box fixing clamp 12, the liquid placing box sliding block 13 is sleeved on a liquid placing box moving sliding rod 15, the liquid placing box moving sliding rod 15 is covered with threads, the liquid placing box moving sliding rod 15 is arranged in the liquid placing box moving rail 16, and the liquid placing box moving rail 16 is fixed above the inner part of the shell 8 through a fixing rod 9.
As shown in fig. 4, the refrigeration module 2 includes a refrigerator 21, a freezing pad 22 and a freezing pad moving rail 23, the refrigerator 21 is connected to the freezing pad 22, and the freezing pad 22 moves left and right through the freezing pad moving rail 23; the refrigerator 21 is a semiconductor refrigerator, an XD-2073 four-pipe double-tower water chiller is selected, a cold water tank and an SC600 cylindrical water pump cold water tank are arranged in the refrigerator, the cold water tank is provided with a refrigerator water inlet 211 and a refrigerator water outlet 212, the freezing pad 22 is provided with a first freezing connector 221 and a second freezing connector 222, the refrigerator water inlet 211 is connected with the first freezing connector 221, the refrigerator water outlet 212 is connected with the second freezing connector 222, one side of the freezing pad 22 is connected with a freezing pad sliding block 225 through a freezing pad fixing clamp 223, the freezing pad sliding block 225 is provided with a second driving motor 226, the freezing pad sliding block 225 is sleeved on a freezing pad moving sliding rod 224, and the freezing pad moving sliding rod 224 is arranged in the freezing pad moving track 23.
The working principle of the water chiller 21 is as follows:
the water cooler comprises radiating fins, a radiating pipe, a fan and a semiconductor refrigerating sheet, wherein the radiating fins are welded with the radiating pipe; the water in the cold water tank is communicated with the water pipe through the water pump and flows through the semiconductor refrigeration sheet and then flows to the position of the refrigeration cushion through the water outlet pipe; the water in the freezing cushion flows back to the cold water tank through the water inlet pipe again for circulating refrigeration.
The water cooler 24 dissipates excessive heat through the heat pipe 242, and the fan 243 and the heat sink 241 accelerate heat dissipation.
As shown in fig. 5-6, the powder pressing module 3 includes a pressing block 31, a stabilizing block 32, a supporting rod 33, an expansion link 34, a pressing moving slider 35 and a pressing moving rail 36, the pressing block 31 is connected to the stabilizing block 32 through a connecting block 321, the stabilizing block 32 is tapered and has a stabilizing effect, the stabilizing block 32 is welded to one end of the supporting rod 33, the other end of the supporting rod 33 is connected to the expansion link 34, the expansion link 34 is a reciprocating electric push rod of MNTL type, the expansion link 34 is connected to the pressing moving slider 35, a third driving motor 351 is disposed on the pressing moving rail 36, the pressing moving slider 35 slides on the pressing moving rail 36, the pressing moving rail 36 is a linear guide rail of TRH20B type, the pressing moving rail 36 is fixed to the lower portion inside the housing 8 through a fixing rod 9, the standby module 5 is arranged below the grinding pressure moving track 36.
As shown in fig. 5 and 7, the dispensing module 4 includes a powder leakage plate 41, a drawer main body 42, a dispensing heat preservation box 43, a brush cleaner 44, a brush support rod 45, a brush slider 46 and a brush slide rod 47, the brush slider 46 is disposed on the brush slide rod 47, the brush slide rod 47 is fixed below the inside of the housing 8 through the support rod 45, two sets of brush sliders 46 and the brush slide rod 47 are disposed, the brush cleaner 44 is disposed therebetween, the brush cleaner 44 moves along with the movement of the brush slider 46, the powder leakage plate 41 is disposed at positions within the four support rods 45, the powder leakage plate 41 is disposed on the drawer main body 42, the dispensing heat preservation box 43 is disposed between the powder leakage plate 41 and the drawer main body 42, and the dispensing heat preservation box 43 is provided with a plurality of partitioned box bodies for containing freeze-dried powder.
As shown in fig. 8, the power module 6 is disposed at one side of the refrigerator 21, the control module 7 is disposed below the refrigerator 21 and the power module 6, the power module 6 is provided with an external port, and the control module 7 is provided with an external port; the model of the power supply module 6 is X-HX0108A, and the model of the control module 7 is CPU224 XP.
As shown in fig. 9-11, a filling opening 82 is disposed at the top of the casing 8, a filling cover 81 is disposed on the filling opening 82, a handle 811 is disposed at the top of the filling cover 81, two locking cover protrusions 812 opposite to each other in position are disposed at the bottom of the filling opening 82, the filling opening 82 is in an inverted cone shape and has an upper opening and a lower opening, a locking cover groove 822 is disposed around the filling opening 82, two locking cover recesses 821 are disposed above the locking cover groove 822, an operation table 83 and a visible window 84 are disposed on the front surface of the casing 8, a casing external opening 86 is disposed on the back surface of the casing, a heat dissipation hole 85 is disposed on one side surface of the casing 8, and a small maintenance window 87 is disposed on the other side surface of the casing.
The console 83 is selected from an XD-W2039 digital control panel.
The first, second and third driving motors 14, 226 and 351 are small round motors of type 1230.
When in use, the cell culture solution is poured from the solution adding port 82, and at the moment, the first half body solution placing box 111 and the second half body solution placing box 112 are in a combined state; then starting the refrigerator 21, enabling the refrigerant liquid to flow through the freezing pad 22 along a pipeline, placing the freezing pad 22 below the liquid placing box 11, after the culture liquid in the liquid placing box 11 is frozen and formed, moving the liquid placing box sliding block 13, enabling the first half liquid placing box 111 and the second half liquid placing box 112 to be changed into an open state, enabling the freeze-dried culture liquid to fall onto the powder leaking plate 41, moving the grinding pressure moving sliding block 35, moving the grinding pressure block 31 above the freeze-dried culture liquid in cooperation with the telescopic rod 34 to perform crushing action, and then enabling the freeze-dried culture liquid to be changed into a freeze-dried powder from a freeze-dried block; and then the grinding block 31 is reset, the cleaning brush 44 is driven by the cleaning brush sliding block 46 to clean the freeze-dried powder into the powder leakage hole 411, and then the freeze-dried powder enters the split charging heat preservation box 43, and the freeze-dried powder is prepared.
The foregoing shows and describes the general principles, essential features, and advantages of the invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, and that the preferred embodiments of the invention and described in the specification are only preferred embodiments of the invention and are not intended to limit the invention, and that various changes and modifications may be made without departing from the novel spirit and scope of the invention, which fall within the scope of the invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (8)
1. A mesenchymal stem cell culture solution freeze-drying process is characterized by comprising the following steps:
s1: screening and selecting umbilical cord donors to carry out pathological detection; taking healthy umbilical cord stem cells to perform umbilical cord mesenchymal stem cell culture, and collecting mesenchymal stem cell conditioned medium;
s2: centrifuging the mesenchymal stem cell conditioned culture solution, filtering the upper suspension, adding a freeze-drying protective agent and glycerol, and uniformly stirring by ultrasonic waves;
s3: putting the obtained liquid into a liquid adding port of a mesenchymal stem cell culture solution freeze-drying device;
s4: waiting for the organism to be subjected to freeze-drying processing to obtain the freeze-dried powder.
2. A mesenchymal stem cell culture solution freeze-drying device is characterized by comprising a solution containing module (1), a refrigerating module (2), a powder pressing module (3), a split charging module (4), a standby module (5), a power supply module (6), a control module (7) and a casing (8); put liquid module (1) set up in refrigeration module (2) top, powder pressing module (3) are located partial shipment module (4) one side for remove extremely partial shipment module (4) top, powder pressing module (3) below is equipped with spare module (5), refrigeration module (2) one side is equipped with power module (6), refrigeration module (2) with power module (6) below is equipped with control module (7).
3. The mesenchymal stem cell culture solution freeze-drying device of claim 2, wherein: the liquid placing module (1) comprises a liquid placing box (11), a liquid placing box fixing clamp (12), a liquid placing box sliding block (13), a first driving motor (14), a liquid placing box moving sliding rod (15) and a liquid placing box moving track (16); the liquid storage box comprises a liquid storage box body (11), wherein two ends of the liquid storage box body (11) are integrally connected with a liquid storage box fixing block (115), the liquid storage box body (11) comprises a first half body liquid storage box (111) and a second half body liquid storage box (112), a half body bulge (113) is arranged at the open end of the first half body liquid storage box (111), a half body recess (114) is arranged at the open end of the second half body liquid storage box (112), the half body bulge (113) is matched with the half body recess (114) in shape, the liquid storage box fixing clamp (12) clamps and fixes the liquid storage box fixing block (115), the liquid storage box sliding block (13) is welded at the bottom of the liquid storage box fixing clamp (12), the liquid storage box sliding block (13) is sleeved on the liquid storage box moving sliding rod (15), the liquid storage box moving sliding rod (15) is covered with threads, and the liquid storage box moving sliding rod (15) is arranged in a liquid storage box moving track (16), the liquid placing box moving track (16) is fixed above the inner part of the machine shell (8) through a fixing rod (9).
4. The mesenchymal stem cell culture solution freeze-drying device of claim 2, wherein: the refrigeration module (2) comprises a refrigerator (21), a freezing pad (22) and a freezing pad moving track (23), the refrigerator (21) is connected with the freezing pad (22), and the freezing pad (22) moves left and right through the freezing pad moving track (23); the refrigerator (21) is a semiconductor refrigerator and is provided with a refrigerator water inlet (211) and a refrigerator water outlet (212), the freezing pad (22) is provided with a first freezing interface (221) and a second freezing interface (222), the refrigerator water inlet (211) is connected with the first freezing interface (221), the refrigerator water outlet (212) is connected with the second freezing interface (222), one side of the freezing pad (22) is connected with a freezing pad sliding block (225) through a freezing pad fixing clamp (223), the freezing pad sliding block (225) is provided with a second driving motor (226), the freezing pad sliding block (225) is sleeved on a freezing pad moving sliding rod (224), and the freezing pad moving sliding rod (224) is arranged in the freezing pad moving track (23).
5. The mesenchymal stem cell culture solution freeze-drying device of claim 2, wherein: the powder pressing module (3) comprises a grinding block (31), a stabilizing block (32), a supporting rod (33), a telescopic rod (34), a grinding pressure moving sliding block (35) and a grinding pressure moving track (36), wherein the grinding block (31) is connected with the stabilizing block (32) through a connecting block (321), the stabilizing block (32) is conical and has a stabilizing effect, the stabilizing block (32) is welded at one end of the supporting rod (33), the other end of the supporting rod (33) is connected with the telescopic rod (34), the telescopic rod (34) is a reciprocating electric push rod of which the model is MNTL, the telescopic rod (34) is connected with the grinding pressure moving sliding block (35), a third driving motor (351) is arranged on the grinding pressure moving track (36), the grinding pressure moving sliding block (35) slides on the grinding pressure moving track (36), and the grinding pressure moving track (36) is fixed below the inner part of the machine shell (8) through a fixing rod (9), the standby module (5) is arranged below the grinding pressure moving track (36).
6. The mesenchymal stem cell culture solution freeze-drying device of claim 2, wherein: the split charging module (4) comprises a powder leakage plate (41), a drawer main body (42), a split charging heat preservation box (43), a sweeping brush (44), a sweeping supporting rod (45), a sweeping sliding block (46) and a sweeping sliding rod (47), wherein the sweeping sliding block (46) is arranged on the sweeping sliding rod (47), the sweeping sliding rod (47) is fixed below the inner part of the shell (8) through the supporting rod (45), the sweeping sliding block (46) and the sweeping sliding rod (47) are provided with two groups, the sweeping brush (44) is arranged between the sweeping sliding block (46), the sweeping brush (44) moves along with the movement of the sweeping sliding block (46), the powder leakage plate (41) is arranged at the position inside the four supporting rods (45), the powder leakage plate (41) is arranged on the drawer main body (42), the heat preservation box (43) is arranged between the powder leakage plate (41) and the drawer main body (42), the split charging heat preservation box (43) is provided with a plurality of separated box bodies for containing freeze-dried powder.
7. The mesenchymal stem cell culture solution freeze-drying device of claim 2, wherein: the refrigerator is characterized in that the power supply module (6) is arranged on one side of the refrigerator (21), the control module (7) is arranged below the refrigerator (21) and the power supply module (6), the power supply module (6) is provided with an external port, and the control module (7) is provided with an external port.
8. The mesenchymal stem cell culture solution freeze-drying device of claim 2, wherein: casing (8) top is equipped with filling opening (82), be equipped with filling opening (81) on filling opening (82), filling opening (81) top is equipped with handle (811), and the bottom is equipped with two relative lockcover arch (812) in position, filling opening (82) are the back taper, upper and lower opening, the round is equipped with lockcover recess (822) above filling opening (82) lockcover recess (822) top sets up two lockcover notches (821), casing (8) openly are equipped with operation panel (83) and visual window (84), and the back is equipped with the outer mouth of casing (86), and the one end side is equipped with louvre (85), and the other end side is equipped with maintenance small window (87).
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