CN115491303A - Mesenchymal stem cell culture device - Google Patents
Mesenchymal stem cell culture device Download PDFInfo
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- CN115491303A CN115491303A CN202211237777.0A CN202211237777A CN115491303A CN 115491303 A CN115491303 A CN 115491303A CN 202211237777 A CN202211237777 A CN 202211237777A CN 115491303 A CN115491303 A CN 115491303A
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- 238000004113 cell culture Methods 0.000 title claims abstract description 26
- 210000002901 mesenchymal stem cell Anatomy 0.000 title claims abstract description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 75
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 claims description 76
- 229910002092 carbon dioxide Inorganic materials 0.000 claims description 38
- 239000001569 carbon dioxide Substances 0.000 claims description 38
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 27
- 229910052760 oxygen Inorganic materials 0.000 claims description 27
- 239000001301 oxygen Substances 0.000 claims description 27
- 239000000243 solution Substances 0.000 claims description 12
- 230000000149 penetrating effect Effects 0.000 claims description 8
- 238000002347 injection Methods 0.000 claims description 6
- 239000007924 injection Substances 0.000 claims description 6
- 230000001939 inductive effect Effects 0.000 claims description 3
- 239000000523 sample Substances 0.000 claims description 3
- 210000000130 stem cell Anatomy 0.000 abstract description 25
- 238000010257 thawing Methods 0.000 abstract description 12
- 210000004027 cell Anatomy 0.000 abstract description 10
- 230000005389 magnetism Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 210000001778 pluripotent stem cell Anatomy 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
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Abstract
The invention relates to the technical field of stem cell culture, in particular to a mesenchymal stem cell culture device, which comprises a box body and a constant-temperature water bath box, wherein the constant-temperature water bath box is fixed at the bottom end in the box body, the upper end of the constant-temperature water bath box is provided with a plurality of test tube grooves, thawing test tubes which extend downwards into the constant-temperature water bath box are inserted in the test tube grooves, the peripheries of the thawing test tubes are fixedly sleeved with limit plates positioned above the constant-temperature water bath box, the middle part of the upper end of the constant-temperature water bath box is rotatably connected with a fixed disc, the left side and the right side of the upper end of the fixed disc are provided with grooves, culture dishes are placed in the grooves, and the lower ends of the culture dishes are connected with magnetic suction plates; according to the invention, the stem cells are placed in the unfreezing test tube, then the unfreezing test tube is inserted into the constant-temperature water bath box, and water is blocked by the unfreezing test tube, so that direct contact between water and the stem cells can be prevented on the premise of normal unfreezing of the cells, and the stem cells are prevented from being polluted due to contact of the water.
Description
Technical Field
The invention relates to the technical field of stem cell culture, in particular to a mesenchymal stem cell culture device.
Background
The mesenchymal stem cell is a pluripotent stem cell, has all commonalities of the stem cell, namely self-renewal and multidirectional differentiation capacity, has extremely wide clinical application prospect, needs to be thawed for the cryopreserved cell in the cell culture process of the mesenchymal stem cell, and needs to be transferred to a proper environment for cell culture after the thawing of the mesenchymal stem cell is completed;
the Chinese patent application No. 202021646840.2 discloses a cell culture device for mesenchymal stem cell culture, which comprises a box body, wherein one side of the box body is hinged with a box door, the bottom of the box body is provided with a constant-temperature water bath, the box body is provided with a cylinder, the cylinder is connected with a motor, the motor is connected with a rotating shaft penetrating into the box body, the rotating shaft is provided with a fixed disk, the fixed disk is fixedly provided with a culture dish through an embracing block, a fixed frame arranged at the top of the box body is arranged above the culture dish, and the fixed frame is provided with a micro-injector penetrating into the box body; the box body is also provided with an oxygen inlet pipe and a carbon dioxide inlet pipe; the cell culture device is simple in structure and convenient to operate, can directly perform subsequent culture on thawed cells through the constant-temperature water bath, can prevent the cells from being polluted in the transfer process, and can adjust and maintain the gas environment required in the cell culture process by adding the structures such as the temperature sensor, the humidity sensor, the carbon dioxide sensor and the like;
however, the invention has the following disadvantages that since the microinjector injects the culture solution into the lower net culture dish, the upper end of the culture dish is open, when the stem cells in the culture dish are thawed, the culture dish is directly placed into the constant temperature water bath, so that the water in the constant temperature water bath may enter the culture dish, the stem cells in the culture dish are easily contacted with the water in the constant temperature water bath directly, and the stem cells are contacted with the water to cause pollution.
Disclosure of Invention
The invention provides a mesenchymal stem cell culture device, which is used for preventing stem cells from being directly contacted by water in a constant-temperature water bath tank in a thawing process, preventing an external environment from interfering with a culture environment in a box body and ensuring normal culture of the cells.
In order to solve the technical problems, the invention adopts the following technical scheme:
a mesenchymal stem cell culture device comprises a box body and a constant temperature water bath box, wherein the constant temperature water bath box is fixed at the bottom end in the box body;
the upper end of the constant temperature water bath box is provided with a plurality of test tube grooves, thawing test tubes which extend downwards into the constant temperature water bath box are inserted into the test tube grooves, and limiting plates which are positioned above the constant temperature water bath box are fixedly sleeved on the peripheries of the thawing test tubes;
the middle part of the upper end of the constant-temperature water bath box is rotatably connected with a fixed disc, the left side and the right side of the upper end of the fixed disc are respectively provided with a groove, culture dishes are respectively placed in the grooves, the lower ends of the culture dishes are respectively connected with a magnetic suction plate, and the bottom ends of the grooves are respectively connected with an electromagnet;
a rotating shaft is fixed in the middle of the upper end of the fixed disc, the rotating shaft penetrates through the box body in an upward rotating mode, a servo motor is fixed at the upper end of the box body, and the upper end of the rotating shaft is connected with a power end of the servo motor;
a culture solution injection assembly is arranged above the culture dish;
an environment control assembly is arranged outside the culture dish.
Furthermore, the left upper end of the constant temperature water bath box is communicated with a water inlet pipe which penetrates out of the box body towards the left, and the left lower end of the constant temperature water bath box is communicated with a water drain pipe which penetrates out of the box body towards the left.
Further, the culture solution injection assembly comprises two mounting plates, microsyringes, a support plate, a pressing plate, a storage battery, a positive and negative motor and a screw rod, the two mounting plates are fixed at the left end and the right end of the rotating shaft respectively, at least two mounting grooves are formed in the upper end of each mounting plate in a penetrating mode, and the microsyringes are located in each mounting groove and are inserted one in each mounting groove.
Furthermore, the support plate is positioned on the periphery of each micro-injector and is fixedly sleeved with one support plate, the support plate is arranged above the mounting plate, and the pressing plate is sleeved on the periphery of the rotating shaft in a sliding manner and is arranged above the micro-injectors.
Further, the storage battery is fixed in the clamp plate upper end, positive and negative motor is fixed in axis of rotation outside upper end, the lead screw upper end connect in positive and negative motor power end, the down screw thread of lead screw runs through the clamp plate, the lead screw lower extreme rotates to be connected in one of them piece the mounting panel upper end.
Further, the environmental control subassembly includes heater, carbon dioxide measuring pump, oxygen measuring pump, carbon dioxide intake pipe, oxygen intake pipe, temperature sensor, humidity transducer and carbon dioxide consistency transmitter, the heater be equipped with two and install respectively in the left and right ends of box inner wall, carbon dioxide measuring pump and oxygen measuring pump down are fixed in from last in proper order the box right-hand member.
Furthermore, the carbon dioxide air inlet pipe and the oxygen air inlet pipe are respectively communicated with the left ends of the carbon dioxide metering pump and the oxygen metering pump, and the carbon dioxide air inlet pipe and the oxygen air inlet pipe penetrate into the box body towards the left.
Further, temperature sensor, humidity transducer and carbon dioxide concentration sensor turn left and be fixed in proper order the box upper end, temperature sensor, humidity transducer and carbon dioxide concentration sensor's inductive probe all down runs through into in the box.
The invention has the beneficial effects that:
1. the stem cells are placed in the thawing test tube, then the thawing test tube is inserted into the constant-temperature water bath box, and water is blocked by the thawing test tube, so that direct contact between water and the stem cells can be prevented on the premise of normal cell thawing, and the stem cells are prevented from being polluted due to contact of water;
2. the magnetic attraction plate is adsorbed by the electromagnet, so that the culture dish can be stably placed in the groove of the fixed disk, the culture dish is prevented from being separated from the fixed disk when rotating, and the uniformity of stem cells in the culture dish during culture can be ensured and the quality of stem cell culture can be ensured when the culture dish rotates;
3. when injecting the culture solution into the culture dish through microinjector, press the microinjector down through the clamp plate to can convenient and fast inject the culture solution in all microinjectors into the culture dish together, and when operating the microinjector, need not to open the box, can avoid external environment to influence the interior culture environment of box, in order to avoid external environment to influence the normal culture of stem cell in the culture dish.
Drawings
FIG. 1 is a schematic view of the overall structure of the present invention;
FIG. 2 is an enlarged partial structural view of the invention A;
FIG. 3 is a schematic view of the structure of the fixing plate of the present invention;
fig. 4 is a schematic diagram of the structure of the mounting plate of the present invention.
Description of reference numerals:
the device comprises a box body 1, a constant-temperature water bath box 2, a water inlet pipe 3, a water outlet pipe 4, a test tube groove 5, a limiting plate 6, a fixed disc 7, a culture dish 8, an electromagnet 9, a magnetic attraction plate 10, a servo motor 11, a rotating shaft 12, a heater 14, a positive and negative motor 15, a screw rod 16, a pressing plate 17, a mounting plate 18, a micro-injector 19, a storage battery 20, a temperature sensor 21, a humidity sensor 22, a carbon dioxide concentration sensor 23, a supporting plate 24, a slot 25, a carbon dioxide metering pump 26, an oxygen metering pump 27, a carbon dioxide inlet pipe 28, an oxygen inlet pipe 29 and a unfreezing test tube 30.
Detailed Description
In order to facilitate understanding of those skilled in the art, the present invention is further described below with reference to the following examples and the accompanying drawings, which are not intended to limit the present invention.
As shown in fig. 1 and 2, in the present embodiment, the device includes a box body 1 and a constant temperature water bath box 2, the constant temperature water bath box 2 is fixed at the bottom end in the box body 1, the upper end of the constant temperature water bath box 2 is provided with a plurality of test tube slots 5, thawing test tubes 30 extending downwards into the constant temperature water bath box 2 are inserted into the test tube slots 5, the peripheries of the thawing test tubes 30 are fixedly sleeved with limit plates 6 positioned above the constant temperature water bath box 2, the left upper end of the constant temperature water bath box 2 is communicated with a water inlet tube 3 penetrating through the box body 1 to the left, and the left lower end of the constant temperature water bath box 2 is communicated with a water outlet tube 4 penetrating through the box body 1 to the left;
at first through inlet tube 3 toward the water that the temperature is suitable of adding in the constant temperature water bath 2, until water fills up constant temperature water bath 2, the water in the constant temperature water bath 2 can contact with the test tube 30 that unfreezes this moment, thereby make the stem cell in the test tube 30 that unfreezes unfreeze, because constant temperature water bath 2 supports in limiting plate 6 below, do benefit to and prevent that the test tube 30 that unfreezes from falling into in constant temperature water bath 2 through test tube groove 5, so that the water in the constant temperature water bath 2 directly produces the contact with the stem cell, consequently, can normally dissolve under the prerequisite of freeze-dried cell, can also prevent that water and stem cell from producing direct contact, so that the stem cell is contacted by water and produces the pollution.
As shown in fig. 1 and 3, in this embodiment, a fixed disk 7 is rotatably connected to the middle of the upper end of the constant temperature water bath tank 2, grooves are formed in the left and right sides of the upper end of the fixed disk 7, culture dishes 8 are placed in the grooves, magnetic attraction plates 10 are connected to the lower ends of the culture dishes 8, electromagnets 9 are connected to the inner bottom ends of the grooves, a rotating shaft 12 is fixed to the middle of the upper end of the fixed disk 7, the rotating shaft 12 is upwardly rotated to penetrate through the tank body 1, a servo motor 11 is fixed to the upper end of the tank body 1, and the upper end of the rotating shaft 12 is connected to a power end of the servo motor 11;
put into the recess on the fixed disk 7 with culture dish 8 in, simultaneously be the power supply of electro-magnet 9 through battery 20, can adsorb the magnetism suction disc 10 of culture dish 8 below after electro-magnet 9 starts, thereby can be with culture dish 8 firm putting on fixed disk 7, will unfreeze test tube 30 again and up take out from test tube groove 5, then will unfreeze the cell after 30 internal unfreezing and pour into culture dish 8, when servo motor 11 circular telegram work and drive axis of rotation 12 and rotate, axis of rotation 12 can drive fixed disk 7 and rotate together, consequently culture dish 8 can be rotatory along with fixed disk 7, because electro-magnet 9 adsorbs with magnetism suction disc 10, thereby do benefit to and produce the separation with fixed disk 7 when preventing culture dish 8 rotatory, and when culture dish 8 is rotatory, do benefit to and make in stem cell evenly distributed to culture dish 8, homogeneity when can guaranteeing culture dish 8 internal stem cell culture, and guarantee the quality of stem cell culture.
As shown in fig. 1, in this embodiment, an environment control assembly is arranged outside the culture dish 8, the environment control assembly includes a heater 14, a carbon dioxide metering pump 26, an oxygen metering pump 27, a carbon dioxide intake pipe 28, an oxygen intake pipe 29, a temperature sensor 21, a humidity sensor 22 and a carbon dioxide concentration sensor 23, the heater 14 is provided with two heaters and is respectively installed at the left end and the right end of the inner wall of the box body 1, the carbon dioxide metering pump 26 and the oxygen metering pump 27 are sequentially fixed at the right end of the box body 1 from top to bottom, the carbon dioxide intake pipe 28 and the oxygen intake pipe 29 are respectively communicated with the left ends of the carbon dioxide metering pump 26 and the oxygen metering pump 27, the carbon dioxide intake pipe 28 and the oxygen intake pipe 29 both penetrate into the box body 1 from left, the temperature sensor 21, the humidity sensor 22 and the carbon dioxide concentration sensor 23 are sequentially fixed at the upper end of the box body 1 from left, and inductive probes of the temperature sensor 21, the humidity sensor 22 and the carbon dioxide concentration sensor 23 all penetrate into the box body 1 from bottom;
in the stem cell culture process in the culture dish 8, the box body 1 is in a closed state, and the water in the constant temperature water bath box 2 is discharged through the drain pipe 4, when the heater 14 is used for heating the interior of the box body 1 to a proper temperature, so as to prevent the water in the constant temperature water bath box 2 from evaporating into the box body 1, and prevent the water in the constant temperature water bath box 2 from causing unnecessary influence on the environment in the box body 1, and an external proper amount of carbon dioxide is introduced into the carbon dioxide inlet pipe 28 through the carbon dioxide metering pump 26, and the carbon dioxide concentration in the box body 1 is detected through the carbon dioxide concentration sensor 23, so that the normal amount of the carbon dioxide in the box body 1 is favorably ensured, and external oxygen is introduced into the oxygen inlet pipe 29 through the oxygen inlet pipe 29, so that the oxygen in the box body 1 is introduced into the box body 1 through the oxygen inlet pipe 29, so as to favorably ensure the oxygen content in the box body 1, and the temperature and the humidity in the box body 1 are detected through the temperature sensor 21 and the humidity sensor 22, so as to favorably control the interior of the box body 1 to culture dry cells under a proper culture environment, thereby ensuring the normal culture of the stem cells.
As shown in fig. 1 and 4, in this embodiment, a culture solution injection assembly is disposed above the culture dish 8, the culture solution injection assembly includes mounting plates 18, micro-syringes 19, support plates 24, a pressure plate 17, a storage battery 20, a positive and negative motor 15, and a screw rod 16, the mounting plates 18 are disposed with two blocks and fixed to the left and right ends of the rotating shaft 12, at least two mounting grooves 25 are penetrated through the upper ends of the mounting plates 18, one micro-syringe 19 is inserted into each mounting groove 25, one support plate 24 is fixedly sleeved on the periphery of each micro-syringe 19, the support plate 24 is disposed above the mounting plate 18, the pressure plate 17 is slidably sleeved on the periphery of the rotating shaft 12 and disposed above the micro-syringes 19, the storage battery 20 is fixed to the upper end of the pressure plate 17, the positive and negative motor 15 is fixed to the upper end of the outer side of the rotating shaft 12, the upper end of the screw rod 16 is connected to the power end of the positive and negative motor 15, the screw rod 16 penetrates through the pressure plate 17 by downward threads, and the lower end of the screw rod 16 is rotatably connected to the upper end of one of the mounting plate 18;
when need inject the culture solution into culture dish 8, under the prerequisite of the culture solution is equipped with in microinjector 19, because mounting panel 18 supports in extension board 24 top, and microinjector 19 is down insert the mounting groove, be the power supply of positive and negative motor 15 through battery 20, consequently start positive and negative motor 15 and drive lead screw 16 and rotate, lead screw 16 can drive clamp plate 17 and move down along axis of rotation 12, thereby make clamp plate 17 down press down all microinjector 19 simultaneously, thereby can convenient and fast inject the culture solution in all microinjector 19 into culture dish 8 together, and during the operation microinjector 19, need not to open box 1, can avoid external environment to influence the interior culture environment of box 1, in order to avoid external environment to influence the normal culture of dry cell in the culture dish 8.
All technical features in the embodiment can be freely combined according to actual needs.
The above embodiments are preferred implementations of the present invention, and other implementations are also included, and any obvious substitutions without departing from the spirit of the present invention are within the scope of the present invention.
Claims (8)
1. The utility model provides a mesenchymal stem cell culture apparatus, includes box (1) and constant temperature water bath (2), constant temperature water bath (2) are fixed in bottom, its characterized in that in box (1):
a plurality of test tube grooves (5) are formed in the upper end of the constant-temperature water bath box (2), unfreezing test tubes (30) which extend downwards into the constant-temperature water bath box (2) are inserted into the test tube grooves (5), and limiting plates (6) which are positioned above the constant-temperature water bath box (2) are fixedly sleeved on the peripheries of the unfreezing test tubes (30);
the middle part of the upper end of the constant-temperature water bath box (2) is rotatably connected with a fixed disc (7), the left side and the right side of the upper end of the fixed disc (7) are respectively provided with a groove, culture dishes (8) are respectively placed in the grooves, the lower ends of the culture dishes (8) are respectively connected with a magnetic suction plate (10), and the bottom ends of the grooves are respectively connected with an electromagnet (9);
a rotating shaft (12) is fixed in the middle of the upper end of the fixed disc (7), the rotating shaft (12) rotates upwards to penetrate through the box body (1), a servo motor (11) is fixed at the upper end of the box body (1), and the upper end of the rotating shaft (12) is connected with a power end of the servo motor (11);
a culture solution injection assembly is arranged above the culture dish (8);
an environment control component is arranged outside the culture dish (8).
2. The mesenchymal stem cell culture apparatus of claim 1, wherein: the upper left end of the constant temperature water bath tank (2) is communicated with a water inlet pipe (3) penetrating out of the tank body (1) to the left, and the lower left end of the constant temperature water bath tank (2) is communicated with a water outlet pipe (4) penetrating out of the tank body (1) to the left.
3. The mesenchymal stem cell culture apparatus of claim 1, wherein: the culture solution injection assembly comprises mounting plates (18), microsyringes (19), support plates (24), a pressing plate (17), a storage battery (20), a positive and negative motor (15) and a screw rod (16), wherein the two mounting plates (18) are respectively fixed at the left end and the right end of the rotating shaft (12), at least two mounting grooves (25) are formed in the upper end of each mounting plate (18) in a penetrating mode, and each microsyringe (19) is located in each mounting groove (25) and is inserted into one mounting groove.
4. The mesenchymal stem cell culture apparatus of claim 3, wherein: the support plate (24) is positioned on the periphery of each micro-injector (19) and is fixedly sleeved with one support plate (24), the support plate (24) is arranged above the mounting plate (18), and the pressing plate (17) is sleeved on the periphery of the rotating shaft (12) in a sliding manner and is arranged above the micro-injectors (19).
5. The mesenchymal stem cell culture apparatus of claim 4, wherein: battery (20) are fixed in clamp plate (17) upper end, positive and negative motor (15) are fixed in axis of rotation (12) outside upper end, lead screw (16) upper end meet in positive and negative motor (15) power end, lead screw (16) down the screw thread runs through clamp plate (17), lead screw (16) lower extreme rotate connect in one of them mounting panel (18) upper end.
6. The mesenchymal stem cell culture apparatus of claim 1, wherein: the environment control assembly comprises a heater (14), a carbon dioxide metering pump (26), an oxygen metering pump (27), a carbon dioxide air inlet pipe (28), an oxygen air inlet pipe (29), a temperature sensor (21), a humidity sensor (22) and a carbon dioxide concentration sensor (23), wherein the heater (14) is provided with two air inlet pipes and two air inlet pipes respectively installed at the left end and the right end of the inner wall of the box body (1), and the carbon dioxide metering pump (26) and the oxygen metering pump (27) are sequentially fixed at the right end of the box body (1) from top to bottom.
7. The mesenchymal stem cell culture apparatus of claim 6, wherein: the carbon dioxide air inlet pipe (28) and the oxygen air inlet pipe (29) are respectively communicated with the left ends of the carbon dioxide metering pump (26) and the oxygen metering pump (27), and the carbon dioxide air inlet pipe (28) and the oxygen air inlet pipe (29) penetrate into the box body (1) leftwards.
8. The mesenchymal stem cell culture apparatus of claim 6, wherein: temperature sensor (21), humidity transducer (22) and carbon dioxide concentration sensor (23) are fixed in proper order toward a left side box (1) upper end, the inductive probe of temperature sensor (21), humidity transducer (22) and carbon dioxide concentration sensor (23) all down runs through into in box (1).
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