CN210620843U - Simulation culture device for stem cells - Google Patents
Simulation culture device for stem cells Download PDFInfo
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- CN210620843U CN210620843U CN201920543323.3U CN201920543323U CN210620843U CN 210620843 U CN210620843 U CN 210620843U CN 201920543323 U CN201920543323 U CN 201920543323U CN 210620843 U CN210620843 U CN 210620843U
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Abstract
The utility model relates to a simulation culture apparatus of stem cell, including stem cell nest, heart district, liver district, lung district, spleen district, kidney district and stomach district, the both ends in stem cell nest are connected with lung district and spleen district respectively, the stomach district for nutrition basin and with the liver district is connected, the heart district is connected with liver district, spleen district and lung district respectively, the spleen district with the kidney district is connected, the kidney district sets up the inlet and the waste discharge mouth, the lung district sets up air inlet and gas vent, set up first peristaltic pump in the heart district, the stem cell nest with set up the third control valve between the spleen district. Compared with the prior art, the utility model provides a pair of stem cell's emulation culture apparatus, the device can guarantee stem cell and can adapt to patient's blood environment fast in the applied link through the culture environment of simulating patient's physiological sign in advance, makes stem cell have abundant quantity and activity and reachs the target point and make the function in parallel.
Description
Technical Field
The utility model belongs to the technical field of stem cell culture, concretely relates to stem cell's emulation culture apparatus.
Background
With the release of the stem cell market in China, the domestic supply of stem cells is seriously insufficient, the traditional 2D culture mode is converted into 3D culture, and the supply quantity of the stem cells can be effectively improved. However, the application of stem cells is not only a matter of the number of stem cells, but also depends on the survival number of stem cells after entering the blood of a patient, and finally depends on the number and activity of stem cells reaching a target. Taking the process of the stem cells entering the blood of a patient as an example, the blood in the blood vessel has systolic pressure and diastolic pressure changes, the calculation is carried out according to the pulse of the human for 60 times/minute, the stem cells entering the blood are stressed by the systolic pressure and the diastolic pressure once per second, the stem cells without pressure-resistant training are difficult to adapt to the blood environment, and the application effect of the stem cells is difficult to effectively ensure.
The stem cell simulation culture method has the advantages that the future application environment (physiological signs of a patient) of the stem cells is simulated, the simulation culture of the stem cells is carried out in vitro, the large-scale amplification and propagation of the stem cells which can adapt to the physiological signs of the patient are guaranteed, the number of living cells and the activity of the cells after the stem cells enter the blood of the patient are greatly improved, and the treatment quality of the stem cells is improved.
SUMMERY OF THE UTILITY MODEL
The utility model provides a stem cell simulation culture device, which can be applied in large-scale industrialization and effectively improve the living cell number and the homing capacity of the stem cells after entering the blood of a patient.
The utility model discloses a design is: culturing stem cells in a stem cell simulation culture device, and simulating the living environment of human stem cells by using a circulating device, such as: guarantee the discharge etc. of temperature, nutrition, acid-base balance, oxygen balance, carbon dioxide balance, metabolite of stem cell culture system, especially make stem cell culture environment and (future application stem cell) patient in vivo stem cell's physiological environment similar through stem cell emulation culture, guarantee living cell number and homing ability behind the stem cell entering patient's blood, promote stem cell treatment, the concrete scheme of the utility model is as follows:
the utility model provides a stem cell's emulation culture apparatus, including stem cell nest, heart district, liver district, lung district, spleen district, kidney district and stomach district, the both ends in stem cell nest are connected with lung district and spleen district respectively, the stomach district for nutrition basin and with the liver district is connected, the heart district is connected with liver district, spleen district and lung district respectively, the stomach district sets up the inlet, the kidney district sets up the inlet and wastes discharge port, the lung district sets up air inlet and gas vent, set up first peristaltic pump in the heart district, the stem cell nest with set up the third control valve between the spleen district.
In any of the above aspects, preferably, a gas permeable membrane is provided in the lung region.
In any of the above aspects, it is preferred that a specific dialysis membrane is provided within the renal region.
In any of the above schemes, preferably, two sets of pipelines are arranged between the kidney region and the spleen region, wherein one pipeline is provided with a fourth control valve, and the other pipeline is provided with a third peristaltic pump.
In any of the above schemes, preferably, a second peristaltic pump is disposed on the air inlet of the lung area, and a second control valve is disposed on the air outlet.
In any of the above schemes, preferably, the stomach area is a replaceable nutrient solution storage tank, the stomach area is connected with power gas, the pressure is kept in a range of 10-60mmHg, and fresh nutrient solution and power for delivering the nutrient solution are supplied to the liver area through the one-way valve at any time.
The utility model also provides a method for the above-mentioned device is used to carry out stem cell simulation culture, and the method includes the following steps according to the sequence:
(1) wrapping the stem cells in gel simulating an extracellular matrix environment for culturing, and protecting the stem cells and regulating the growth of the stem cells by using the gel;
(2) arranging the stem cells wrapped by the gel in a stem cell nest, arranging the stem cell nest in a constant-temperature area, wherein the culture temperature of the area is consistent with the body temperature of a patient to which the stem cells are applied, the characteristics of the constant-temperature area are consistent with the characteristics of a life field of the patient, nutrient solution in the stem cell nest flows in a single direction, pressure change exists in the flowing process, the changed pressure characteristics and frequency are set to be consistent with the characteristics of blood pressure and pulse of the patient to which the stem cells are applied, and the stem cell nest is connected with a circulating device for simulating blood circulation of a human body;
(3) the heart area of the circulating device is utilized to simulate the heart function of a human body to complete the circulation of nutrient solution in the stem cell culture process, and the stem cells have sufficient nutrient supply and discharge metabolic waste through the circulation of the nutrient solution; the lung area of the circulating device is utilized to simulate the function of the human lung, the gas exchange of nutrient solution is completed through a specific gas permeation membrane, and the balance of oxygen and carbon dioxide in the stem cell culture process is kept; the spleen area of the circulating device is utilized to simulate the functions of the spleen of a human body to finish the temporary storage of the nutrient solution, so that the nutrient solution enters the nutrient solution again for circulation after metabolic wastes are removed; the kidney area of the circulating device is utilized to simulate the function of the kidney of a human body, and metabolic waste of stem cells is removed through a specific dialysis membrane device, so that the stem cells are protected from being inhibited by the metabolic waste; the liver area of the circulating device is utilized to simulate the liver function of a human body, and nutrient solution is obtained from a nutrient solution storage tank (stomach area) to complete the supplement and supply of the nutrient solution of the circulating system;
(4) various physiological parameters and life field parameters of a patient with stem cells applied in the future are effectively acquired through a control platform connected with the circulating device, the whole stem cell culture system is controlled to achieve a specific culture environment for simulating physiological signs of the patient, and stem cell simulation culture is carried out to obtain the stem cell.
Preferably, the stem cells are wrapped by calcium alginate gel in cooperation with extracellular matrix substances, the throttle valve is matched with the heart area to simulate the change of systolic pressure and diastolic pressure of a human body and pulse, oxygen supply is enhanced by regulating the oxygen partial pressure of the lung area, metabolic waste in the stem cell culture process is effectively discharged through the kidney, and a stem cell culture environment simulating physiological signs of a patient is formed by coordinating a stem cell nest and a stem cell culture auxiliary system through the cooperation of a control platform and cloud data.
The specific area of stem cell culture is specially called stem cell nest, the stem cell nest completes the simulation culture of stem cells by means of the circulating device and the control platform, the circulating device is composed of a plurality of functional partitions, and each functional partition is respectively specially called according to the functional characteristics: the heart area, the lung area, the spleen area, the kidney area and the liver area (including the stomach area) can simulate the functional organs of a human body to provide nutrition for the growth of stem cells and discharge metabolic waste, and the functions of the specific parts are as follows:
the stem cell culture process of the utility model uses a device simulating the human heart function to complete the circulation of nutrient solution, and the stem cells have sufficient nutrient supply and discharge metabolic waste through the circulation of the nutrient solution, and the part is specially called as the heart area; in the stem cell culture process, a device which simulates the function of the lung of a human body and is composed of a specific gas permeation membrane is used for completing the gas exchange of nutrient solution, and the balance of oxygen and carbon dioxide in the stem cell culture process is kept, wherein the part is specially called a lung area; in the stem cell culture process, a device simulating the functions of the spleen of a human body is used for finishing the temporary storage of nutrient solution, the part is specially called the spleen area, and the nutrient solution in the spleen area enters the nutrient solution circulation again after metabolic waste is removed; the device for simulating the kidney function of a human body is used for discharging the metabolic waste of the stem cells in the stem cell culture process, the part is specially called as a kidney area, and the kidney area is provided with a semipermeable membrane device which can effectively remove the metabolic waste generated in the stem cell growth process; the stem cell culture process uses a device for simulating the functions of human liver to complete the supplement and supply of nutrient solution, the part is specially called a liver area, the liver area obtains nutrient solution from a nutrient solution storage tank, the nutrient solution storage tank is specially called a stomach area, and nutrient substances are continuously obtained from the outside by replacing the nutrient solution storage tank.
The container and the related components of the pipeline used in the stem cell culture process are disposable, so that the safety of the stem cells is ensured; the related components of the container and the pipeline are made of polypropylene, polystyrene or polyethylene and the like which meet the medical and sanitary requirements, and are sterilized and provided in an aseptic packaging mode; the details are not repeated in the following description.
Stem cell niche: the pressure container consists of a cylinder structure and a cover, wherein an opening is arranged below the cylinder and is connected with a luer connector, so that the pressure container can be assembled quickly; the cover completes the closing of the cylinder through threads, an opening is arranged on the cover and is connected with a luer connector, and a sealable opening is arranged on the cover and is used for assembling the sensor; the outlet of the stem cell nest is provided with a control valve which can be matched with a peristaltic pump of the heart area to generate blood pressure change and pulse characteristics of a simulated human body; meanwhile, the stem cell nest is arranged in a constant temperature and electromagnetic shielding area, and a biological electromagnetic field is added, so that the stem cells can be cultured in a simulation way in an environment simulating the physiological characteristics of a patient to be applied in the future.
The heart area: the peristaltic pump mainly comprises a peristaltic pump and a three-way component, wherein the three-way component is connected with a spleen area, a liver area and a peristaltic pump head, and a culture solution from the spleen area and the liver area can enter a silicone tube in the peristaltic pump head through the three-way component and enter a lung area through pressurization of the peristaltic pump; the peristaltic pump is provided with a multi-roller pump head (such as rollers), the nutrient solution conveying stability is improved, the peristaltic pump is subjected to electromagnetic shielding treatment, and the pump head is used for completing nutrient solution conveying by using a high-quality sterile silicone tube.
Liver region: the pressure control range is 10-60mmHg (measured according to blood pressure standard) for providing culture solution for heart region and spleen region, the liver region is connected with the three-way component of the heart region through a control valve, and the flow is controlled by the control valve to make fresh nutrient solution enter the spleen region or mixed with nutrient solution from the spleen region in different proportions in the three-way component and then enter the heart region; the liver area is also provided with a one-way valve communicated with the stomach area, and the nutrient solution flows to the liver area from the stomach area in one way.
Lung area: the hollow fiber membrane component part is a pressure container and is used as an artificial lung in medical treatment, or is a sterile hollow fiber membrane component meeting medical and health standards, the core of the membrane component is a hollow fiber membrane made of a specific gas permeable membrane (oxygen/carbon dioxide permeable membrane), and the shell of the membrane component is made of materials such as polypropylene, polystyrene or polyethylene; the pressurized nutrient solution flows in the hollow fiber tube, the nutrient solution in the lung area is subjected to gas exchange with air/carbon dioxide mixed gas through the gas permeable membrane and enters the stem cell nest after reaching the set oxygen content/carbon dioxide content parameter, and nutrition and oxygen are provided for the stem cells. The pressure sterile air (containing carbon dioxide) is matched with a control valve, the exchange rate of oxygen and carbon dioxide in the nutrient solution is regulated and controlled by regulating the total gas pressure, and the total amount of oxygen and carbon dioxide in the nutrient solution comprises a dissolved state part and a combined state part. The lung area is provided with two nutrient solution interfaces which are respectively connected with the heart area and the stem cell nest; two gas ports are provided, which are respectively connected with a pressure sterile air (containing carbon dioxide) supply pipeline and a gas discharge pipeline.
Spleen area: the device is a pressure-free container, a cylinder structure or a square barrel structure, is communicated with the outside through a sterile 0.2 micron filter membrane, and is provided with two liquid level sensors; when the liquid level is lower than the lowest set value, the valve in the liver area is opened, and nutrient solution is actively supplied to the spleen area; when the liquid level is higher than the highest set value, the permeation rate of the kidney area is increased. The spleen area is provided with an interface connected with the stem cell nest and used for receiving nutrient solution after stem cell culture; the spleen area is provided with two interfaces which are communicated with the kidney area and used for dialysis discharge of the stem cell metabolic waste in the nutrient solution; the spleen area also has a port connected with the three-way component of the heart area.
Renal region: the pressure container is a hollow fiber membrane component part used for renal dialysis in medical treatment or a sterile hollow fiber membrane component meeting medical and health standards, the core of the membrane component is a hollow fiber membrane made of a specific dialysis membrane, and the shell of the membrane component is made of materials such as polypropylene, polystyrene or polyethylene. The nutrient solution containing the metabolic waste of the stem cells in the spleen area enters the kidney area after being pressurized, the metabolic waste of the stem cells can pass through the dialysis membrane and enter the dialysate under the action of pressure, and the stem cell culture solution without the metabolic waste of the stem cells returns to the spleen area for reuse through a pipeline with a control valve. The kidney area is provided with two nutrient solution interfaces which are connected with the spleen area, and two dialysate interfaces which are used for dialyzing and separating the metabolic wastes of the stem cells from the kidney area. The nutrient solution circulation in the kidney area is adjusted by the cooperation of a peristaltic pump and a control valve to regulate the dialysis rate; the dialysate is disposable and not circulated, but there is also a peristaltic pump and control valves to coordinate the adjustment of the dialysis rate.
Gastric region: the pressure control range is 10-60mmHg (measured according to blood pressure standard) in a pressure container, a cylinder structure or a square barrel structure, and the nutrient solution storage tank can be replaced, so that nutrient solution and nutrient solution conveying power can be provided for a liver region, and the requirement of a stem cell culture process on the nutrient solution is ensured; the stomach area is provided with two interfaces, one interface is connected with the liver area through a one-way valve and supplies culture solution to the liver area in a one-way mode, and the other interface is connected with a sterile power gas pipeline, so that the pressure range of 10-60mmHg is maintained, and nutrient solution and conveying power are supplied to the liver area at any time.
The utility model discloses a growth environment (extracellular matrix) of stem cell makes stem cell grow under the most suitable culture condition in the simulation human body, includes wherein: the gel medium is utilized to simulate the extracellular matrix to provide nutrition and a protection effect for the growth of stem cells, the nutrient solution simulates the unidirectional flow of blood to provide nutrition and discharge metabolic waste for the stem cells, the pressure change is realized in the culture process, the pressure characteristic and frequency are consistent with the blood pressure and pulse characteristics of a patient using the stem cells, the culture temperature of a stem cell nest is consistent with the internal temperature of the patient using the stem cells, the stem cell nest is positioned in a specific life field consistent with the patient using the stem cells, the stem cells can adapt to the blood environment (such as blood pressure) after entering the blood of the human body through simulation culture, and then the stem cells can be transported and homed along with the blood flow.
In addition, the method carries out stem cell culture by simulating human organ functions to maximally approach the characteristics of stem cell epimatrix in human body outside the human body, and is not only suitable for various stem cells, such as: mesenchymal stem cells, etc., and may also be applied to other types of cells, such as: iPS cells, T cells, and the like.
Compared with the prior art, the utility model provides a pair of emulation culture apparatus of stem cell through the culture environment of simulating patient's physiological sign in advance, guarantees that the stem cell can adapt to patient's blood environment fast in the application link, makes the stem cell have abundant quantity and active target site and makes the function in parallel.
Drawings
FIG. 1: schematic diagram of a simulation culture device of stem cells.
The figures are labeled as follows: 1-stem cell nest, 2-heart region, 3-liver region, 4-lung region, 5-spleen region, 6-kidney region, 7-stomach region, 8-first control valve, 9-second peristaltic pump, 10-second control valve, 11-third control valve, 12-third peristaltic pump, 13-fourth control valve, 14-dialysate, 15-fourth peristaltic pump, 16-fifth control valve, 17-gas permeable membrane, 18-specific dialysis membrane, 19-cloud data center, 20-control platform, 21-life field, 22-constant temperature system, 23-sensor, 24-data interface, 25-sterile air, 26-fresh nutrient solution, 27-metabolic waste.
Detailed Description
In order to further understand the contents of the present invention, the present invention will be described in detail with reference to the specific embodiments, if the chemical reagents and instruments used in the present invention are not specially described, they can be purchased from commercial channels, all the following operations can be completed in the local a-level clean area, all the articles are treated by aseptic or bacteriostatic treatment, and they will not be described separately.
Example one
As shown in fig. 1, the present invention provides a stem cell simulation culture apparatus, which mainly comprises the following steps:
s1, according to the composition characteristics of the stem cell extracellular matrix, selecting stem cell living environment substances such as collagen and glycan to be fully mixed with the gel with high biocompatibility and wrap the gel on the periphery of the stem cell, so as to provide protection for the subsequent stem cell culture and provide nutrition for the stem cell growth and discharge metabolic waste;
s2, loading the stem cells protected by the gel into a stem cell nest 1, and starting a circulating device (the assembly and culture parameter setting are completed in advance) to culture the stem cells, wherein the specific process is as follows:
(1) before the device is operated, the spleen area 5 is free of nutrient solution, the first control valve 8 of the liver area 3 is opened, the stomach area 7 actively conveys fresh nutrient solution 26 to the spleen area 5 and reaches the lowest set value of the liquid level of the spleen area 5, under the power driving of the heart area (first peristaltic pump) 2, the nutrient solution reaches the heart area (first peristaltic pump) 2 and enters the lung area 4, and the nutrient solution receives oxygen and carbon dioxide from the gas permeable membrane 17 in the lung area 4 and then enters the stem cell nest 1 to provide nutrition and oxygen for the stem cells wrapped in the simulated extracellular matrix; gas exchange is powered by the cooperation of a second peristaltic pump 9 and a second control valve 10, oxygen and carbon dioxide are provided by sterile air 25;
(2) the nutrient solution flows out of the stem cell nest 1 through a third control valve 11 and enters the spleen area 5;
(3) part of nutrient solution in the spleen area 5 and fresh nutrient solution 26 from the liver area 3 enter a heart area (a first peristaltic pump) 2 together, and then enter a stem cell nest 1 after passing through a lung area 4 to supply nutrition and oxygen for stem cells; the other part is driven by a third peristaltic pump 12 to enter the renal region 6 and return to the splenic region 5 through a fourth control valve 13; the kidney area is provided with the specific dialysis membrane 18 which can filter metabolic waste 27 such as urea and the like generated in the growth process of the stem cells, so that the metabolic waste 27 is prevented from influencing the growth of the stem cells; the dialysate 14 enters the kidney region 6 through a fourth peristaltic pump 15 and is discharged through a fifth control valve 16;
(4) the control platform 20 can acquire relevant instructions from the cloud data center 19 to control stem cell culture, and meanwhile, upload stem cell culture data to the cloud data center 19; the control platform 20 is also responsible for providing a patient life field 21 for stem cell growth for future use of stem cells; the culture temperature of the system is maintained by a constant temperature system 22, the system state is monitored by a sensor 23, and the system is connected with a control platform 20 through a data interface 24.
The utility model discloses circulating device who uses, including drawing stem cell nest 1, heart district 2, liver district 3, lung district 4, spleen district 5, kidney district 6 and stomach district 7, stem cell nest 1's both ends are connected with lung district 4 and spleen district 5 respectively, stomach district 7 with liver district 3 is connected, heart district 2 is connected with liver district 3, spleen district 5 and lung district 4 respectively, stomach district 7 sets up the inlet, kidney district 6 sets up the inlet and the mouth of wasting discharge, lung district 4 sets up air inlet and gas vent, set up first peristaltic pump in the heart district 2, set up third control valve 11 between stem cell nest 1 and the spleen district 5.
In any of the above embodiments, it is preferable that a gas permeable membrane 17 is provided in the lung region 4.
In any of the above solutions it is preferred that a specific dialysis membrane 18 is provided within the renal area 6.
In any of the above schemes, two sets of pipelines are preferably arranged between the renal area 6 and the spleen area 5, wherein one pipeline is provided with a fourth control valve 13, and the other pipeline is provided with a third peristaltic pump 12.
In any of the above solutions, it is preferable that the inlet of the lung area 4 is provided with a second peristaltic pump 9, and the outlet is provided with a second control valve 10.
In any of the above embodiments, it is preferable that the stomach region 7 is a replaceable nutrient solution storage tank, and the stomach region 7 is connected with a sterile power gas, and the pressure is maintained in the range of 10-60mmHg, so as to provide fresh nutrient solution 26 and power for delivering nutrient solution for the liver region 3 through a one-way valve at any time.
Example two
S1, according to the composition characteristics of the growing environment of the stem cells, selecting collagen, fibronectin, laminin and hyaluronic acid to mix with the stem cells, wrapping the stem cell mixed solution with 1.5% sodium alginate glue solution, forming solidified calcium alginate gel by using 100mmol/L calcium ions and wrapping the gel on the periphery of the stem cells, so as to protect the stem cells and ensure that the stem cells can effectively take in nutrition and discharge metabolic waste in the growing process;
s2, loading the stem cells protected by the gel into a stem cell nest 1, and starting a circulating device (the assembly and culture parameter setting are completed in advance) to culture the stem cells, wherein the specific process is as follows:
(1) before the device is operated, the spleen area 5 is free of nutrient solution, the first control valve 8 of the liver area 3 is opened, the stomach area 7 actively conveys fresh nutrient solution 26 to the spleen area 5 and reaches the lowest set value of the liquid level of the spleen area 5, under the power driving of the heart area (first peristaltic pump) 2, the nutrient solution reaches the heart area (first peristaltic pump) 2 and enters the lung area 4, and the nutrient solution receives oxygen and carbon dioxide from the gas permeable membrane 17 in the lung area 4 and then enters the stem cell nest 1 to provide nutrition and oxygen for the stem cells wrapped in the simulated extracellular matrix; gas exchange is powered by the cooperation of a second peristaltic pump 9 and a second control valve 10, oxygen and carbon dioxide are provided by sterile air 25;
(2) the nutrient solution flows out of the stem cell nest 1 through a third control valve 11 and enters the spleen area 5;
(3) part of nutrient solution in the spleen area 5 and fresh nutrient solution 26 from the liver area 3 enter a heart area (a first peristaltic pump) 2 together, and then enter a stem cell nest 1 after passing through a lung area 4 to supply nutrition and oxygen for stem cells; the other part is driven by a third peristaltic pump 12 to enter the renal region 6 and return to the splenic region 5 through a fourth control valve 13; the kidney area is provided with the specific dialysis membrane 18 which can filter metabolic waste 27 such as urea and the like generated in the growth process of the stem cells, so that the metabolic waste 27 is prevented from influencing the growth of the stem cells; the dialysate 14 passes through a fourth peristaltic pump 15 to the renal region 6 and is discharged through a fifth control valve 16;
(4) the control platform 20 can acquire relevant instructions from the cloud data center 19 to control stem cell culture, and meanwhile, upload stem cell culture data to the cloud data center 19; the control platform 20 is also responsible for providing a patient life field 21 for stem cell growth for future use of stem cells; the culture temperature of the system is maintained by a constant temperature system 22, the system state is monitored by a sensor 23, and the system is connected with a control platform 20 through a data interface 24;
(5) through human body function simulation, stem cells can grow in the physiological sign environment of a patient who applies the stem cells in the future, and the cultured stem cells can be subjected to the hammer training of the change of systolic pressure and diastolic pressure and the change of pulse of the blood pressure of the patient who applies the stem cells in the future, so that the stem cells have better cell survival rate after entering the blood of the patient, and the homing capability and efficacy are guaranteed to be realized.
Compared with the prior art
TABLE 1 Effect of culture methods on Stem cell culture
A existing 2D culture technique: pressure-free culture, pH7.35, temperature 36.5 deg.C, 5% carbon dioxide content, standard culture solution, cell survival rate of 85%, blood survival rate of 35%, and homing ability of 5%.
B existing 2D culture technology: pressure-free culture, pH7.45, temperature of 37.5 deg.C, 20% carbon dioxide content, standard culture solution, cell survival rate of 80%, blood survival rate of 33%, and homing ability of 0.5%.
C existing 2D culture techniques: pressure-free culture, pH7.40, temperature of 37 ℃, carbon dioxide content of 10 percent, standard culture solution, cell survival rate of 85 percent, blood survival rate of 33 percent and homing capacity of 3 percent.
D, simulation culture technology: the cell survival rate is 90%, the blood survival rate is 45% and the homing capacity is 3% when the pressure is 100mmHg, the pH value is 7.35, the temperature is 36.5 ℃, the oxygen content (including dissolved oxygen and bound oxygen) is 10mL/100mL of culture solution, the carbon dioxide content (including dissolved state and bound state) is 55mL/100mL of culture solution and the simulated pulse is 60 times/min.
E, simulation culture technology: the cell survival rate is 95%, the blood survival rate is 50% and the homing capacity is 11% when the artificial pulse is 80 times/min under the conditions that the pressure is 120mmHg, the pH value is 7.40, the temperature is 37.0 ℃, the oxygen content (including dissolved oxygen and bound oxygen) is 20mL/100mL of culture solution, the carbon dioxide content (including dissolved state and bound state) is 45mL/100mL of culture solution and the simulated pulse is 80 times/min.
F, simulation culture technology: the cell survival rate is 90%, the blood survival rate is 45% and the homing capacity is 5% when the artificial pulse is 100 times/min under the conditions that the pressure is 160mmHg, the pH value is 7.45, the temperature is 37.5 ℃, the oxygen content (including dissolved oxygen and bound oxygen) is 10mL/100mL of culture solution, the carbon dioxide content (including dissolved state and bound state) is 55mL/100mL of culture solution and the simulated pulse is 100 times/min.
G simulation culture technology: the cell survival rate is 93 percent, the blood survival rate is 49 percent and the homing capacity is 7 percent when the artificial pulse is performed for 70 times/min under the conditions that the pressure is 180mmHg, the pH value is 7.35, the temperature is 37.5 ℃, the oxygen content (including dissolved oxygen and bound oxygen) is 20mL/100mL of culture solution, the carbon dioxide content (including dissolved state and bound state) is 55mL/100mL of culture solution and the simulated pulse is 70 times/min.
H, simulation culture technology: the cell survival rate is 90%, the blood survival rate is 45% and the homing capacity is 6% when the artificial pulse is 60 times/min under the conditions that the pressure is 120mmHg, the pH value is 7.35, the temperature is 36.5 ℃, the oxygen content (including dissolved oxygen and bound oxygen) is 10mL/100mL of culture solution, the carbon dioxide content (including dissolved state and bound state) is 55mL/100mL of culture solution and the simulated pulse is 60 times/min.
As shown in table 1, compared with the prior art, the stem cells cultured by the 3D simulation culture method of the present invention have higher culture survival rate, survival rate of entering blood and homing ability than the prior 2D culture technique.
It will be understood by those skilled in the art that the apparatus for simulated culture of stem cells of the present invention includes any combination of the above-mentioned aspects and embodiments of the present invention and the parts shown in the drawings, which is limited to space and is not described in any combination for the sake of brevity. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (6)
1. The utility model provides a simulation culture apparatus of stem cell, its characterized in that, includes stem cell nest, heart district, liver district, lung district, spleen district, kidney district and stomach district, the both ends in stem cell nest are connected with lung district and spleen district respectively, the stomach district for nutrition stock tank and with the liver district is connected, the heart district is connected with liver district, spleen district and lung district respectively, the spleen district with the kidney district is connected, the kidney district sets up the inlet and the waste discharge mouth, the lung district sets up air inlet and gas vent, set up first peristaltic pump in the heart district, the stem cell nest with set up the third control valve between the spleen district.
2. The simulated stem cell culture device of claim 1, wherein a gas permeable membrane is disposed in the lung region.
3. The simulated culture device of stem cells as claimed in claim 1, wherein a specific dialysis membrane is arranged in the kidney region.
4. The apparatus for simulating stem cell culture according to claim 1, wherein two sets of pipelines are disposed between the renal region and the splenic region, wherein a fourth control valve is disposed on one of the pipelines, and a third peristaltic pump is disposed on the other pipeline.
5. The apparatus for simulating stem cell culture according to claim 1, wherein a second peristaltic pump is disposed on the air inlet of the lung region, and a second control valve is disposed on the air outlet.
6. The simulated stem cell culture device of claim 1, wherein the gastric region is coupled to a motive gas.
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| CN113755432A (en) * | 2020-07-17 | 2021-12-07 | 上海我武干细胞科技有限公司 | Stem cell culture method |
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