Disclosure of Invention
The invention aims to provide a stem cell culture perfusion device, wherein a layer of heat insulation pipe is sleeved outside a hose, a plurality of rollers on a roller wheel can be firstly contacted with the heat insulation pipe and then extrude the hose through the heat insulation pipe, so that a first contact object of the rollers is not the hose any more and is limited by the reason that the heat insulation pipe is made of an aerogel hose or a rubber and plastic hose, the heat conductivity coefficient is reduced, the probability of temperature rise of stem cell solution in the hose due to long-time extrusion is reduced, the survival time of the finally cultured stem cells is prolonged, and the increment quantity is increased, and the problems in the background technology are solved.
In order to achieve the purpose, the invention provides the following technical scheme:
a stem cell culture perfusion device, comprising:
the peristaltic pump comprises a driving unit, a pump head and a hose, wherein the pump head consists of an upper cover and a base, rollers are rotatably mounted in the base and consist of a plurality of rotating rollers, the output end of the driving unit is connected with the rollers, and the hose is placed on the rollers of the base;
further comprising:
prevent the intensification subassembly, be equipped with in the pump head and be used for preventing the subassembly that elevates temperature that prevents that the hose heaies up in transmission process.
Preferably, prevent rising temperature the subassembly including placing the thermal-insulated pipe in the pump head, and thermal-insulated pipe and hose laminating, the aerogel hose or the rubber and plastic hose of microminiature bore can be selected to the material of thermal-insulated pipe.
When stem cells are perfused, a peristaltic pump is generally used for being matched with a hose to perfuse stem cell solution, a roller is arranged in a pump head of the peristaltic pump and consists of a plurality of rollers, the rollers are matched with the inner wall of the pump head and are used for uniformly rotating and extruding the hose, a pump with a larger pillow volume is formed between the two rollers, and the part of the stem cell solution in the hose can be extruded forwards and moved after being uniformly rotated in cycles to form regular perfusion;
however, the hose is circularly extruded by a plurality of rollers for a long time, the temperature of the extruded part of the hose is increased due to the heat generated by long-time extrusion friction, and then the temperature is transmitted to the stem cell solution in the hose through the hose, so that the temperature of the stem cell solution is likely to rise when the stem cell solution passes through the hose in the pump head, and the temperature has certain influence on the stem cell culture environment, for example, the isolated and purified mouse spermatogonial stem cells are cultured in vitro at the temperature of 30, 34 and 37 degrees, and the experimental result shows that the average survival time of the cells at 34 degrees is longest, the increment quantity is the greatest, and the rise of the temperature is likely to cause the reduction of the average survival time of the cells and the increment quantity;
the heat insulation pipe is arranged in the pump head and is hollow, the heat insulation pipe can be sleeved outside the hose, when the hose is extruded and discharged by the plurality of rollers on the roller, the rollers can be firstly contacted with the heat insulation pipe and then extrude the hose through the heat insulation pipe, so that a first contact object of the plurality of rollers is not the hose any more and is limited by the material reason of the heat insulation pipe, the temperature transfer coefficient is increased, the probability of temperature rise of stem cell solution in the hose due to long-time extrusion is reduced, the survival time of the finally cultured stem cells is prolonged, and the increment quantity is increased; most hoses are disposable consumables, the heat insulation pipes can be used for a long time, and the material value of the heat insulation pipes can be relatively high, so that the stability of long-term operation is guaranteed.
The thermal conductivity coefficient of the aerogel is very low, 0.018W/(K.m) at normal temperature, even can reach 0.009W/(K.m) at low temperature, while the thermal conductivity coefficient of the PVC hose is 0.034W/(K.m) at normal temperature, the material of the thermal insulation pipe is an aerogel hose or a rubber and plastic hose, the thermal insulation pipe is sleeved outside the hose, the first object in contact with the rotating rod on the roller is converted into the thermal insulation pipe on the frictional extrusion with the roller, and the thermal conductivity coefficient is very low because the thermal insulation pipe is made of the aerogel hose, so that the heat generated by the friction between the roller and the thermal insulation pipe can be basically only attached to the outer surface of the thermal insulation pipe and cannot permeate into the hose inside of the thermal insulation pipe, and the situation that the dry cell solution in the hose is heated due to long-term friction of the roller during perfusion of the peristaltic pump is ensured to a certain extent, the survival rate of the cultured dry cells is reduced, and the increment quantity is small is caused.
Preferably, the heat insulation pipe comprises two semicircular half pipes, two half pipes are respectively installed on the upper cover and the base, two inserting rods are arranged on the pipe wall of one side of each half pipe, inserting grooves matched with the corresponding inserting rods are formed in the pipe walls of the other sides of the two half pipes, and each inserting groove is a bevel groove.
The heat insulation pipe is arranged in the pump head, when the hose needs to be matched with the roller, the heat insulation pipe is required to be sleeved outside the hose, the heat insulation pipe is arranged to be composed of two half pipes, the two branch pipes are respectively installed on the upper cover and the base, due to the matching of the insertion rod and the insertion groove, an experimenter only needs to open the upper cover, the hose is placed in the half pipe on the base, the upper cover is closed again, each insertion groove is a bevel groove, the insertion rod can be inserted into the corresponding insertion groove, the two half pipes can be smoothly clamped, the hose can be wrapped, the heat insulation pipe is composed of the two half pipes, the two half pipes are respectively arranged in the upper cover and the base, the insertion groove and the insertion rod are matched, the experimenter does not need to add other steps, only needs to place the hose in the half pipe on the base, the upper cover is closed again, the installation of the heat insulation pipe can be completed, the heat insulation pipe can be convenient and fast, the heat insulation pipe is sleeved outside the hose, the preparation action before perfusion is shortened, and the perfusion efficiency is improved.
Preferably, the insulating tube and the hose are in an interference fit.
The interference fit is that the elasticity of the material is utilized to enlarge the hole, the hole is deformed and sleeved on the shaft, the clamping force of the shaft is generated when the hole is restored, two parts are connected, and on the same principle, the thermal insulation pipe is matched with the upper cover and the base through the inserted rod and the slot to realize the interference fit, the upper cover and the base give two half-pipe pressures, the inserted rod and the slot are matched together again, so that the thermal insulation pipe wraps the hose, in a normal state, the diameter of the inner ring of the thermal insulation pipe is equal to the diameter of the outer ring of the hose, the thermal insulation pipe can be better attached to the hose, the interference fit is utilized, the thermal insulation pipe can generate the clamping force of the hose, the gap between the thermal insulation pipe and the hose can be reduced, the relative friction condition is reduced, and when the peristaltic pump runs, the condition that the temperature rise due to the generation of heat can be reduced.
Preferably, a plurality of protruding ribs for extruding the hose are uniformly arranged on the inner wall of the heat insulation pipe, and the shape of each protruding rib is a long strip with a semicircular cross section.
Set up the one deck thermal-insulated pipe between gyro wheel and hose, just can extrude the stem cell solution in the hose forward and move through the thermal-insulated pipe of extrusion at the gyro wheel, thickness has increased, then two adjacent change "pillow" that form between the roller can the widen in two, then the efficiency that the extrusion is advanced has reduced, evenly be equipped with a plurality of protruding muscle on the thermal-insulated pipe inner wall, when the thermal-insulated pipe of gyro wheel extrusion, by the protruding muscle extrusion hose of extrusion department, form a plurality of small spaces, can reduce the space widen of "pillow" body volume both sides, then two change the stem cell solution that the extrusion is advanced between the roller and increase, the efficiency that the extrusion is advanced has improved.
Preferably, an extrusion groove is formed in the upper cover, an extrusion strip for enabling the heat insulation pipe to be attached to the hose is installed in the extrusion groove, and the extrusion strip is a long strip made of silicon rubber.
The heat insulation pipe is sleeved outside the hose, and cannot move relative to the hose, if relative movement occurs, friction still exists between the heat insulation pipe and the hose, and further the hose may still generate temperature rise of stem cell solution in the hose due to heat generation caused by friction, an extrusion groove is formed in the upper cover, an extrusion strip is installed in the extrusion groove and is matched with the extrusion strip, the extrusion strip is made of soft material and has certain ductility and extensibility, when the upper cover is buckled with the base, the extrusion groove can be filled up by the extrusion strip under the action of the elasticity of the extrusion groove and the extrusion strip, and then a half pipe corresponding to the heat insulation pipe is extruded towards the direction of the hose, so that the friction force between the hose and the inner wall of the heat insulation pipe is increased, the probability that the heat insulation pipe and the hose generate friction due to relative movement when the roller operates is reduced, the temperature rise of the hose due to friction is avoided, and further the temperature rise of the stem cell solution in the hose is further, so that the average survival time of cells is shortened, and the number of the increased value is reduced.
Preferably, the edges of the inner rings of the two half pipes are provided with compensation spaces for compensating the extrusion clearance, and the cross sections of the compensation spaces are chamfers.
When two half pipes are extruded and attached, if the edges between the two half pipes are completely attached, then when the two half pipes are extruded, the extrusion force can extrude the two half pipes towards the outside of the half pipes, the similar elongated effect of the two half pipes can be realized, the two half pipes can not be completely attached to the hose, when the roller is in contact friction with the roller, friction can be generated between the half pipes and the hose, the inner ring edges of the two half pipes are provided with compensation spaces, the cross sections of the compensation spaces are chamfers, namely, the gaps of the two chamfers are reserved for compensation, when the two half pipes are extruded, due to the gap reasons existing in the two chamfers, the extruding direction is changed, the half pipes are extruded and filled towards the chamfers, the hose is more attached to the two half pipes and the hose under the effect of the extrusion grooves and the chamfers at the moment, the phenomenon that the heat insulation pipes and the hose generate relative friction is avoided, the hose is caused to appear due to the condition of friction heating, and the stability of the temperature in the hose during perfusion of the peristaltic pump is improved.
Preferably, the two sides in the extrusion groove are provided with wedge blocks for guiding the attachment direction of the extrusion strip.
The extrusion groove is matched with the upper extrusion strip, the extrusion strip can fill the extrusion groove under the action of the elasticity of the extrusion strip, then, the half pipe corresponding to the heat insulation pipe is extruded towards the hose direction, but the extrusion direction has certain disorder, and then the extrusion strip can overflow towards the left end and the right end when extruding the corresponding half pipe, wedge blocks are arranged on two sides inside the extrusion groove, the wedge blocks play a certain guiding role, after the upper cover is closed, the extrusion strip is limited by the extrusion groove, after the extrusion groove is filled, the extrusion strip is extruded towards the corresponding half pipe, under the guiding role of the wedge blocks, the extrusion strip is guided towards the center, the extrusion strip extrudes the corresponding half pipe towards the center, and the half pipe is better attached to the hose.
Preferably, the cross section of the heat insulation pipe is divided into a major arc shape and a minor arc shape.
The cross section of the heat insulation pipe has two states, one state is an arc shape, the arc shape is that one half pipe wraps about 3/4 of the diameter of the outer circle of the hose under the action of the extrusion strip and the compensation chamfer, the other half pipe wraps the rest 1/4 of the outer circle of the hose, and the top half pipe wraps most of the hoses; one kind is inferior arcuation, 1/2 of hose is all wrapped up to upper and lower half pipe promptly, and the stability of parcel does not have the good of major arc, but when the gyro wheel roll extrusion, because the half pipe respectively accounts for 1/2, the extrusion force that the protruding muscle on the half pipe gave the hose is relatively steady, then the stem cell solution of pumping is comparatively steady, plays positive effect to stem cell solution follow-up culture.
Preferably, the base is provided with a through hole, a heat conducting fin for conducting heat is arranged in the through hole, and the heat conducting fin is attached to a half pipe on the base.
After the upper cover and the base lock of pump head, almost form an inclosed cavity in the pump head, the gyro wheel is at the in-process of long-time operation in-process with the friction of heat insulating tube, be aerogel hose or rubber and plastic hose because of the heat insulating tube material, the heat that the friction produced can be piled up on the heat insulating tube surface, can produce certain heat and then the temperature in the airtight space in the pump head can rise, and the produced heat of gyro wheel most all piles up in the gyro wheel both sides, avoid piling up the stem cell solution that leads to in the hose and heaies up because of heat under the airtight environment, set up the through-hole at the upper cover, and be equipped with the conducting strip in the through-hole, and the conducting strip extends to outside the base, then the produced heat of gyro wheel and heat insulating tube friction discharges to the base outside through the conducting strip, reduce the above-mentioned condition.
Compared with the prior art, the invention has the beneficial effects that:
1. according to the stem cell culture perfusion device, the heat insulation pipe is sleeved outside the hose, the rollers can be in contact with the heat insulation pipe firstly, then the hose is extruded through the heat insulation pipe, so that a first contact object of the rollers is not the hose any more and is limited by the reason that the heat insulation pipe is made of an aerogel hose or a rubber and plastic hose, the heat conductivity coefficient is reduced, the probability of temperature rise of stem cell solution in the hose due to long-time extrusion is reduced, the survival time of the finally cultured stem cells is prolonged, and the increment quantity is increased.
2. According to the stem cell culture perfusion device, the heat insulation pipe is arranged and comprises two half pipes, the two half pipes are respectively arranged in the upper cover and the base, and the insertion groove is matched with the insertion rod, so that a laboratory worker does not need to add other steps, only the hose is put into the half pipe on the base, and then the upper cover is closed, the heat insulation pipe can be installed, the heat insulation pipe can be conveniently and quickly sleeved outside the hose, the time consumed by preparation actions before perfusion is reduced, and the perfusion efficiency is improved.
3. According to the stem cell culture perfusion device, the extrusion strip is matched with the two half pipes, the extrusion strip can fill the extrusion groove under the action of the elasticity of the extrusion groove and the extrusion strip, and then the half pipes corresponding to the heat insulation pipes are extruded towards the direction of the hose, so that the friction force between the hose and the inner wall of the heat insulation pipe is increased, and the probability of friction caused by relative displacement between the heat insulation pipe and the hose when the roller operates is reduced; then, through the compensation space, the cross section of the compensation space is a chamfer, a gap between two chamfers is reserved for compensation, when the two half pipes are extruded to be attached, the extrusion direction is changed due to the gap between the two chamfers, the half pipes are extruded and filled towards the chamfer, and at the moment, the two half pipes and the hose are attached to the hose under the action of the extrusion groove and the chamfer; then through wedge and extrusion groove cooperation, the wedge plays certain guide effect, gives the direction of extrusion strip towards center department, and the extrusion strip is towards center department's extrusion to corresponding half pipe for half pipe better with the hose laminating.
Detailed Description
Referring to fig. 1 to 7, the present invention provides a stem cell culture perfusion apparatus, which comprises the following technical schemes:
the first embodiment is as follows:
conditions are as follows: the radiuses of the two half pipes 501 are not consistent, and the radius of the upper half pipe 501 is larger than that of the lower half pipe 501;
the working process comprises the following steps: when culturing stem cells with a large influence of temperature, such as isolated and purified mouse spermatogonial stem cells, the temperature is preferably 34 degrees, and the average survival time of the cells is reduced and the number of proliferation is reduced at 30 degrees and 37 degrees compared with 34 degrees, the influence of the temperature of the solution is also large during culturing.
Before carrying out the perfusion, need put into pump head 2 hose 3 among the perfusion use, the upper cover is opened to the pull rod that the pulling was covered, hose 3 takes out afterwards, put into half pipe 501 on the base in the suitable position of hose 3, the pull rod of pulling back on the upper cover again, the upper cover descends and the base lock, two half pipe 501 block under the effect of inserted bar 502 and slot 503, because of a plurality of slots 503 all are the bevel connection groove, then in the insert slot 503 that a plurality of inserted bars 502 can be smooth, accomplish the preparation work before the perfusion, start peristaltic pump 1 afterwards, begin the perfusion.
The peristaltic pump 1 is started to work, the output end of the driving unit drives the roller 4 to rotate, a plurality of rollers on the roller 4 are all in contact with the heat insulation pipe 5, and the convex ribs 504 in the heat insulation pipe 5 are matched with the inner wall of the upper cover to drive stem cell liquid in the hose 3 to extrude and move forwards and reciprocate circularly; in the process, because the diameter of the upper half pipe 501 is larger than that of the lower half pipe 501, the heat insulation pipe 5 and the hose 3 are in a major arc state at the moment, the extrusion strip 7 extrudes the upper half pipe 501 on the upper cover towards the hose 3 under the action of the elasticity of the extrusion groove 6 and the extrusion strip 7, meanwhile, two wedge blocks 8 in the extrusion groove 6 play a role in guiding, after the upper cover is closed, the extrusion strip 7 is subjected to extrusion force limited by the extrusion groove 6, after the extrusion groove 6 is filled with the extrusion strip, the extrusion strip extrudes towards the corresponding half pipe 501, under the guiding action of the wedge blocks 8, the extrusion strip 7 guides the extrusion strip 7 towards the center, and the extrusion strip 7 extrudes the corresponding half pipe 501 towards the center, so that the half pipe 501 is better attached to the hose 3; because the chamfers 5011 are arranged on the inner ring edges of the two half pipes 501, the gap between the two chamfers 5011 is reserved for compensation, when the two half pipes 501 are extruded to be attached, the extruding direction is changed due to the gap between the two chamfers 5011, the half pipes 501 are extruded and filled towards the chamfers 5011, and then the two half pipes 501 and the hose 3 are attached to the hose 3 under the action of the extruding groove 6 and the chamfers 5011, so that the friction force between the hose 3 and the inner walls of the heat insulation pipes is increased, and the probability of friction caused by relative displacement between the heat insulation pipes 5 and the hose 3 when the roller 4 operates is reduced.
Example two:
conditions are as follows: when the radii of the two half pipes 501 are consistent;
the working process is as follows: before carrying out the perfusion, need put into pump head 2 to hose 3 in the perfusion use, the upper cover is opened to the pull rod that the pulling was covered, take out hose 3 afterwards, put into half pipe 501 on the base in the suitable position of hose 3, the pull rod that again comes back pulling upper cover, the upper cover descends and the base lock, two half pipe 501 block under the effect of inserted bar 502 and slot 503, because of a plurality of slots 503 all are the bevel connection groove, then in a plurality of inserted bar 502 can be smooth insert slot 503, accomplish the preparation work before the perfusion, start peristaltic pump 1 afterwards, begin the perfusion.
The peristaltic pump 1 starts to work, the output end of the driving unit drives the roller 4 to rotate, a plurality of rollers on the roller 4 are all in contact with the heat insulation pipe 5, and the convex ribs 504 inside the heat insulation pipe 5 are matched with the inner wall of the upper cover to drive stem cell liquid in the hose 3 to move forwards in an extruding manner and reciprocate cyclically.
In the process, because the radiuses of the two half pipes 501 are consistent, the space between the heat insulation pipe 5 and the hose 3 is in a minor arc state, the half pipes 501 occupy 1/2 of each other in the minor arc state, the extrusion force given to the hose 3 by the protruding ribs 504 on the half pipes 501 is relatively stable, the pumped stem cell solution is relatively stable, and a positive effect is achieved on the subsequent culture of the stem cell solution, the extrusion strip 7 extrudes the upper half pipe 501 on the upper cover towards the direction of the hose 3 under the self-elasticity effect of the extrusion groove 6 and the extrusion strip 7, meanwhile, the two wedge blocks 8 in the extrusion groove 6 play a guiding role, after the upper cover is closed, the extrusion strip 7 is limited by the extrusion force of the extrusion groove 6, after the extrusion groove 6 is filled, the extrusion strip extrudes towards the corresponding half pipe 501, and under the guiding effect of the wedge blocks 8, the extrusion strip 7 guides the extrusion strip 7 towards the center of the corresponding half pipe 501, so that the half pipe 501 can be better attached to the hose 3; because the chamfers 5011 are arranged on the edges of the inner rings of the two half pipes 501, the gap between the two chamfers 5011 is reserved for compensation, when the two half pipes 501 are extruded to be attached, the extruding direction is changed due to the gap between the two chamfers 5011, the half pipes 501 are extruded and filled towards the chamfers 5011, at this time, the two half pipes 501 and the hose 3 are attached to the hose 3 under the action of the extruding groove 6 and the chamfers 5011, the friction force between the hose 3 and the inner walls of the heat insulation pipes is increased, and the probability that the heat insulation pipes 5 and the hose 3 generate friction due to relative displacement when the roller 4 operates is reduced.