CN111996117B - Placenta blood extraction device and method integrating transportation and collection - Google Patents
Placenta blood extraction device and method integrating transportation and collection Download PDFInfo
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Abstract
The invention discloses a placenta blood extraction device and method integrating transportation and collection, belongs to the technical field of biology, and aims to solve the problem that the pollution probability is high because the number of times of placenta exposure cannot be effectively reduced in the transportation and collection processes of the existing placenta blood. The invention comprises a box body, an upper cover, a lifting platform and a control unit; the lifting platform is arranged in the box body and moves up and down along the inner wall of the box body under the control of the control unit, the lifting platform divides the interior of the box body into an upper area and a lower area, and the upper area is used for loading preservation solution for soaking placenta; the upper surface of the lifting platform is provided with a plurality of spurt tissues for puncturing the placenta to obtain placenta blood when the lifting platform upwards presses the placenta; the upper cover lock closes the upper shed department at the box, the upper cover has the sample connection for connect outside sampling device and collect the placenta blood. The method is used for transporting the placenta and extracting the placenta blood.
Description
Technical Field
The invention relates to a transportation device of placenta, which is a device with the function of collecting placenta blood and reducing pollution probability, and belongs to the technical field of biology.
Background
Placental-derived hematopoietic stem cells are a group of primitive hematopoietic cells present in placental tissue, and hematopoietic stem cells are progenitors of blood cells such as erythrocytes, leukocytes, platelets, and the like, which are highly undifferentiated cells, and are primitive cells of all blood cells, most of which are immune cells.
The content of the hematopoietic stem cells in the placenta tissue is 8-10 times of that in the umbilical cord blood, so that the placenta tissue can be used by children for several times and can even provide treatment for a plurality of adult patients. The placenta hematopoietic stem cell transplantation can effectively solve the technical problems that the source of bone marrow or mobilized peripheral blood is insufficient, the number of the hematopoietic stem cells in umbilical cord blood is not enough for adults, and the like, and is expected to replace the bone marrow, the mobilized peripheral blood and the umbilical cord blood to be used for the hematopoietic stem cell transplantation of allogeneic or syngeneic children.
Transplantation can be used for treating acute leukemia, chronic leukemia, myelodysplastic syndrome, hematopoietic stem cell disease, myeloproliferative disease, lymphoproliferative disease, macrophage disease, hereditary metabolic disease, histiocytic disease, hereditary erythrocyte disease, hereditary immune system disease, hereditary platelet disease, plasma cell disease, thalassemia, non-hematologic malignancy, acute radiation disease, etc. with 75 kinds of lethal diseases such as malignant hematological disease, partial malignancy, partial hereditary disease, etc.
The extraction process for placental blood is roughly: the placenta is filled into a sterile bag, the bag is placed in a box body with an ice bag and transported to a sterile laboratory, the sterile bag with the placenta is taken out from the box body under the sterile environment, the sterile bag is manually squeezed to discharge the placenta blood, and then the placenta blood is collected by devices such as a blood collection bag and the like and is subjected to subsequent conventional operations such as separation, extraction and the like to remove red blood cells and redundant blood plasma, so that the nucleated cell layer rich in hematopoietic stem cells is obtained.
In the traditional placental blood extraction process, the box body with the ice bag can only be used for transportation, and the placenta needs to be taken out after the placenta is transported to a destination, so that the exposure times are increased, and the pollution probability of the placenta is increased.
Disclosure of Invention
The invention aims to solve the problem that the pollution probability is high because the times of placenta exposure cannot be effectively reduced in the transportation and collection processes of the conventional placental blood, and provides a transportation and collection integrated placental blood extraction device and method.
The invention relates to a placenta blood extraction device integrating transportation and collection, which comprises a box body 1, an upper cover 2, a lifting platform 3 and a control unit 8;
the lifting platform 3 is arranged in the box body 1 and moves up and down along the inner wall of the box body 1 under the control of the control unit 8,
the lifting platform 3 divides the interior of the box body 1 into an upper area and a lower area, and the upper area is used for loading preservation solution for soaking placenta;
the upper surface of the lifting platform 3 is provided with a plurality of spurt tissues 4 for puncturing the placenta to obtain placenta blood when the lifting platform 3 upwards presses the placenta;
the upper cover 2 is buckled at the upper opening of the box body 1, and the upper cover 2 is provided with a sampling port 7 for connecting an external sampling device to collect placental blood.
Preferably, the control unit 8 comprises a processor 801, a temperature sensor 802, a refrigeration unit 803 and a communication module 805;
the processor 801 communicates with the mobile phone APP through the communication module 805;
the processor 801 receives a mobile phone APP instruction to control the refrigeration unit 803 to work, and the processor 801 receives a mobile phone APP instruction to control the lifting platform 3 to work;
Preferably, the control unit 8 further includes a position module 804, the position module 804 is configured to monitor information about a position of the box 1, and the processor 801 sends the position signal to the mobile phone APP.
Preferably, the heat insulation layer 6 is further included, and the heat insulation layer 6 is arranged on the inner wall of the box body 1.
Preferably, the temperature monitoring device further comprises a display screen 9, wherein the display screen 9 is arranged on the upper surface of the upper cover 2 and is used for displaying temperature information of the box body 1.
Preferably, the utility model further comprises a handle 10, and the handle 10 is arranged on the upper surface of the upper cover 2.
Preferably, the box body further comprises an observation window 5, wherein the observation window 5 is arranged on the side wall of the box body 1 and is positioned at the upper area of the box body 1.
Preferably, a filter screen is also included, which is arranged at the inlet of the sampling port 7.
Preferably, the upper cover 2 is an automatic lifting cover, and is controlled and completed by the processor 801 according to an instruction issued by the mobile phone APP.
The invention also provides another scheme: a method for extracting placental blood comprises the following steps:
step one, placing the placenta into a preservation solution of a box body 1, and closing an upper cover 2;
secondly, transporting the sample to a destination laboratory in a constant temperature state;
step three, starting a lifting platform 3 in the biological safety cabinet to lift upwards to press the placenta, and puncturing the placenta by the spurt tissue 4 on the lifting platform 3 to enable blood in the placenta to flow out;
step four, along with the pressure applied to the placenta by the lifting platform 3, the blood extruded from the placenta and the preservation solution are discharged into a sampling device through a sampling port 7 together, and the extraction of the placenta blood is completed;
the method is realized based on the placenta blood extracting device which integrates transportation and collection as one body and is set forth in any one of claims 1 to 9.
The invention has the beneficial effects that: the device provides a constant-temperature storage environment for placenta transportation, provides position information for receiving personnel in real time, ensures that the placenta is in a closed state in the whole process to extract the placenta blood without taking out the placenta after the placenta is transported to a laboratory, ensures the transportation temperature and stability of a sample and reduces intermediate steps, improves the survival rate of the obtained placenta blood, increases the number of cloned colonies, reduces the preparation time, relatively increases the expression of the flow-type phenotype CD34 hematopoietic stem cells, controls the closed state pollution rate to be infinitely close to 0% in the whole process from the collection to the preparation, and has the indexes superior to the traditional process in all aspects of the collected placenta blood.
Drawings
Fig. 1 is a schematic structural diagram of a placenta blood extracting device integrating transportation and collection;
FIG. 2 is a control schematic block diagram of the present invention;
FIG. 3 is a graph comparing cloning using the conventional technique and the present technique, in which (a) is a graph showing cloning efficiency in the conventional technique and (b) is a graph showing cloning efficiency in the present technique.
Detailed Description
The first specific implementation way is as follows: the present embodiment will be described with reference to fig. 1 to 3, and the placental blood extraction device integrated with transportation and collection according to the present embodiment includes a box 1, an upper cover 2, a lifting platform 3, and a control unit 8;
the lifting platform 3 is arranged in the box body 1 and moves up and down along the inner wall of the box body 1 under the control of the control unit 8,
the lifting platform 3 divides the interior of the box body 1 into an upper area and a lower area, and the upper area is used for loading preservation solution for soaking placenta; and the lower area is a space for mechanical parts such as a compression bar supporting the operation of the lift table 3.
The upper surface of the lifting platform 3 is provided with a plurality of spurt tissues 4 for puncturing the placenta to obtain placenta blood when the lifting platform 3 upwards presses the placenta;
The control unit 8 includes a processor 801, a temperature sensor 802, a refrigeration unit 803, and a communication module 805;
the processor 801 communicates with the mobile phone APP through the communication module 805;
the processor 801 receives a mobile phone APP instruction to control the refrigeration unit 803 to work, and the processor 801 receives the mobile phone APP instruction to control the lifting platform 3 to work;
The inner wall of the box body 1 is provided with a heat preservation layer 6.
The inlet of the sampling port 7 is provided with a filter screen, and the selected filter screen is a 100-mesh filter screen. The sampling port 7 is connected with an external collecting tube and the like and is used for collecting extruded placental blood, and the placental blood and the preservation solution are mixed together.
The upper cover 2 is an automatic lifting cover and is controlled and completed by the processor 801 according to instructions given by the mobile phone APP.
The device provided by the embodiment can not only complete the transportation of the placenta, but also complete the extraction of the placental blood, reduces the step of taking the placenta out of the transportation box body, and directly completes the extraction of the placental blood in the box body 1, thereby greatly reducing the probability of the contamination of the placenta and completing the collection and extraction of the placental blood in a sterile environment.
The device is held by a worker to obtain the placenta from a placenta source and then is transported to a laboratory. In the transportation process, receiving personnel in a laboratory can obtain the position information of the device in real time through a mobile phone APP, and the time of arriving at the laboratory is estimated, so that the receiving preparation can be made in time; meanwhile, the receiving personnel can also acquire the internal temperature of the box body 1 in real time through the mobile phone APP, and can remotely adjust the constant temperature value to effectively intervene so as to ensure the good preservation environment of the placenta.
The opening and closing of the upper cover 2, the lifting of the lifting platform 3 and the refrigeration of the refrigeration unit 803 are all given by the processor 801, and the instruction is obtained by two ways, one is to arrange corresponding buttons on the extraction device of the embodiment, such as the upper cover 2, which is completed by the staff responsible for transporting the placenta; the other is obtained from the mobile phone APP through the communication module 805, which is done by the staff responsible for transporting the placenta and/or the receiving staff of the laboratory.
The second embodiment is as follows: in the following, the present embodiment is described with reference to fig. 2, and the present embodiment further describes the first embodiment, the control unit 8 further includes a position module 804, the position module 804 is configured to monitor the position information of the box 1, and the processor 801 sends the position signal to the mobile phone APP.
The third concrete implementation mode: the present embodiment will be described with reference to fig. 2, and the present embodiment further describes the first embodiment, and further includes an observation window 5, where the observation window 5 is provided on a side wall of the case 1 and is located in an upper region of the case 1.
The observation window 5 is provided in the present embodiment in order to observe the state of the placenta in the casing 1.
The fourth concrete implementation mode is as follows: the present embodiment will be described with reference to fig. 2, and the method for extracting placental blood according to the present embodiment is implemented by the placental blood extraction device integrated with transportation and collection according to the first or second embodiment, and includes the steps of:
step one, placing the placenta into a preservation solution of a box body 1, and closing an upper cover 2;
the preservation solution in the embodiment is heparin sodium injection with 500ml of electrolyte and 5000IU, and the function of the preservation solution is to ensure the activity and anticoagulation of the sample.
Cleaning the placenta before placing in a preservation solution: washing placenta surface with normal saline for 2-3 times.
Then, the placenta is quickly placed in the case 1 to allow the placenta to be immersed in the preservation solution, and the upper cover 2 is closed.
Secondly, transporting the sample to a destination laboratory in a constant temperature state;
in the transportation mode, the refrigeration unit 803 is activated to maintain a temperature suitable for storing the placenta in the case 1. On sending position information in real time to the cell-phone APP of receiving personnel in the laboratory in the transportation, conveniently do the preparation work.
Step three, starting a lifting platform 3 in the biological safety cabinet to lift upwards to press the placenta, and puncturing the placenta by a spurt tissue 4 on the lifting platform 3 to make blood in the placenta flow out;
after the device reaches a laboratory, the whole device is put into a biological safety cabinet for placenta blood collection.
The lifting platform 3 is started to move upwards, and the stabbing tissues 4 on the lifting platform pierce the placenta to enable the blood in the placenta to flow out and be mixed with the preservation solution.
Step four, along with the pressure applied to the placenta by the lifting platform 3, the blood extruded from the placenta and the preservation solution are discharged into a sampling device through a sampling port 7 together, and the extraction of the placenta blood is completed;
the sampling device comprises 1 collecting three connected bags, 3 syringes of 5ml, 1 syringe of 20ml, 1 syringe of 50ml, 1 single freezing bag, 1 HES hydroxyethyl starch bag, 1-2 freezing tubes of 2ml and 2ml EP tubes. The sampling tube for collecting the three bags is connected with the sampling port 7 of the device, a 100-mesh filter screen is arranged in the sampling port, blood flows into the transfer bag through the sampling port 7, the lifting table 3 fully extrudes the blood in the placenta and then extracts the blood, and in the process of extracting the blood in the placenta, the whole placenta is in a closed state and is not in contact with the outside, so that the possibility of pollution is reduced.
And then carrying out a sample separation process: extracting 1ml of sample in a transfer bag for counting, detecting the WBC number and the RBC volume, if the volume is more than 22ml, adding hydroxyethyl starch according to 20 percent of the total volume of the sample, uniformly mixing for 10min by a shaking table, landing a refrigerated centrifuge for 30-100g, 6-9min and 10 ℃ for centrifugation, smoothly taking out the bag after centrifugation, hanging the bag on a stainless steel hook, slowly discharging the red blood cells at the bottom layer by placing the red blood cell bag on an electronic balance, discharging the red blood cell volume = the total volume of the red blood cells of-22 ml, paying attention to scales on the balance, covering a pipe clamp after red blood discharge, continuing centrifugation for 600-900g, 10-15min and 10 ℃, placing the plasma bag on the electronic balance after centrifugation, discharging blood plasma = sample total volume-22 ml-discharging red blood cell volume, extracting 14ml of blood plasma respectively, injecting 6ml of oxygen and 8ml of anaerobic bottle for aseptic detection, remaining 21ml of sample, taking 1ml for inspection for counting, survival rate, phenotype and cloning, clamping the rest with 4 ℃ ice bags, putting into a 4 ℃ refrigerator for precooling for 15min, adding final concentration 50% DMSO and 5% dextran self-prepared cryopreservation solution according to the proportion of 1.
The traditional method and the method of the invention are respectively used for 15 comparison tests, and the method of the invention is used for collection and preparation, so that the transportation temperature and the stability of the sample are ensured, the intermediate steps of the sample are reduced, and the survival rate is greatly improved compared with the traditional method, and the method is shown in table 1.
TABLE 1 comparison of Activity ratios
Sequence of | Percent activity of the traditional method | Activity value of the method of the invention% |
1 | 92.4 | 99.8 |
2 | 92.5 | 99.0 |
3 | 87.7 | 98.1 |
4 | 93.0 | 99.6 |
5 | 94.1 | 98.3 |
6 | 92.3 | 98.5 |
7 | 90.3 | 97.9 |
8 | 93.5 | 99.2 |
9 | 96.7 | 99.3 |
10 | 97.2 | 99.7 |
11 | 89.8 | 98.5 |
12 | 93.1 | 97.8 |
13 | 93.1 | 99.5 |
14 | 95.7 | 99.7 |
15 | 93.9 | 98.3 |
Mean value of | 93.0±1.2 | 98.8±0.8 |
The results of 15 comparative tests carried out by the conventional method and the method of the present invention respectively show that the placenta blood is more fully collected by the extrusion of the device of the present invention, and sufficient nucleated cell number is obtained, and the comparison with the nucleated cell number of the conventional method is shown in table 2.
TABLE 2 comparison of nucleated cell numbers
Sequence of | |
Number of cells in the method of the |
1 | 16.2 | 30.2 |
2 | 15.5 | 25.7 |
3 | 19.5 | 19.9 |
4 | 16.0 | 21.3 |
5 | 20.3 | 25.0 |
6 | 19.7 | 26.7 |
7 | 16.6 | 19.6 |
8 | 22.3 | 20.2 |
9 | 22.5 | 22.6 |
10 | 18.7 | 23.4 |
11 | 16.2 | 28.8 |
12 | 18.5 | 26.8 |
13 | 16.1 | 26.9 |
14 | 13.5 | 18.7 |
15 | 16.5 | 19.6 |
Average number | 17.9±1.1 | 23.7±0.6 |
Using 15 comparative experiments with each of the conventional and inventive methods, the number of colony clones obtained was significantly greater for the inventive method than shown in Table 3. A comparison of the two is shown in figure 3.
TABLE 3 comparison of colony clones
Sequence of | Colony cloning by traditional method | The method of the present invention colonies several clones |
1 | 15 | 29 |
2 | 20 | 33 |
3 | 13 | 31 |
4 | 22 | 32 |
5 | 21 | 36 |
6 | 23 | 28 |
7 | 13 | 33 |
8 | 14 | 35 |
9 | 18 | 35 |
10 | 20 | 28 |
11 | 19 | 26 |
12 | 18 | 38 |
13 | 26 | 30 |
14 | 27 | 34 |
15 | 30 | 31 |
Mean number of | 19.9±0.8 | 31.9±0.5 |
By performing 15 comparative tests on each of the conventional method and the method of the present invention, the preparation time of the present invention is saved as compared to the prior art, which is specifically shown in table 4.
TABLE 4 preparation time comparison
Sequence of | The preparation time of the traditional method is short | The preparation time of the method is short |
1 | 2.2 | 1.5 |
2 | 2.1 | 1.5 |
3 | 2.3 | 1.6 |
4 | 2.0 | 2.0 |
5 | 2.0 | 2.0 |
6 | 2.9 | 1.6 |
7 | 2.6 | 2.1 |
8 | 2.1 | 1.9 |
9 | 2.2 | 1.8 |
10 | 2.2 | 2.0 |
11 | 2.0 | 2.0 |
12 | 2.3 | 2.0 |
13 | 2.0 | 1.5 |
14 | 2.1 | 1.3 |
15 | 2.2 | 1.3 |
Average number | 2.2±1.3 | 1.7±0.4 |
In 15 comparative experiments using the conventional method and the method of the present invention, the expression of the flowing phenotype CD34 hematopoietic stem cells is relatively increased, which is shown in Table 5.
TABLE 5 comparison of expression of flow-phenotype CD34 hematopoietic Stem cells
Sequence of | Conventional methods for phenotypic assay% | The method of the invention tests the phenotype% |
1 | 0.43 | 0.56 |
2 | 0.49 | 0.59 |
3 | 0.41 | 0.54 |
4 | 0.39 | 0.60 |
5 | 0.42 | 0.49 |
6 | 0.41 | 0.48 |
7 | 0.43 | 0.55 |
8 | 0.44 | 0.54 |
9 | 0.38 | 0.58 |
10 | 0.44 | 0.52 |
11 | 0.42 | 0.51 |
12 | 0.36 | 0.56 |
13 | 0.47 | 0.57 |
14 | 0.49 | 0.58 |
15 | 0.36 | 0.49 |
Average number | 0.42±0.04 | 0.54±0.02 |
The whole process from collection to preparation is closed state pollution rate control infinitely close to 0%, and all indexes of the collected placental blood are superior to those of the traditional process.
Claims (10)
1. The placenta blood extraction device integrating transportation and collection is characterized by comprising a box body (1), an upper cover (2), a lifting platform (3) and a control unit (8);
the lifting platform (3) is arranged in the box body (1) and moves up and down along the inner wall of the box body (1) under the control of the control unit (8),
the lifting platform (3) divides the interior of the box body (1) into an upper area and a lower area, and the upper area is used for loading preservation solution for soaking placenta;
the upper surface of the lifting platform (3) is provided with a plurality of spurt tissues (4) which are used for puncturing the placenta to obtain placenta blood when the lifting platform (3) upwards presses the placenta;
upper cover (2) lock is at the upper shed department of box (1), upper cover (2) have sample connection (7) for connect outside sampling device and collect placental blood.
2. The placenta blood extraction device of claim 1, wherein the control unit (8) comprises a processor (801), a temperature sensor (802), a refrigeration unit (803), and a communication module (805);
the processor (801) communicates with the mobile phone APP through the communication module (805);
the processor (801) receives a mobile phone APP instruction to control the refrigeration unit (803) to work, and the processor (801) receives the mobile phone APP instruction to control the lifting platform (3) to work;
temperature sensor (802) are used for monitoring box (1) inside temperature, and processor (801) send this temperature information to cell-phone APP.
3. The placenta blood extraction device integrating transportation and collection according to claim 2, wherein the control unit (8) further comprises a position module (804), the position module (804) is used for monitoring position information of the box body (1), and the processor (801) sends the position signal to the mobile phone APP.
4. The placenta blood extraction device of claim 1, which integrates transportation and collection, further comprising a heat insulation layer (6), wherein the heat insulation layer (6) is disposed on the inner wall of the box body (1).
5. The placenta blood extraction device integrating transportation and collection according to claim 1, further comprising a display screen (9), wherein the display screen (9) is disposed on the upper surface of the upper cover (2) and used for displaying temperature information of the box body (1).
6. The placenta blood extraction device with integrated transportation and collection of claim 1, further comprising a handle (10), wherein the handle (10) is disposed on the upper surface of the upper cover (2).
7. The placenta blood extraction device with integrated transportation and collection of claim 1, further comprising an observation window (5), wherein the observation window (5) is disposed on a side wall of the box body (1) and is located in an upper region of the box body (1).
8. The placenta blood extraction device of claim 1, which integrates transportation and collection, further comprising a filter screen disposed at the inlet of the sampling port (7).
9. The placenta blood extraction device with integrated transportation and collection of claim 1, wherein the upper cover (2) is an automatic lifting cover and is controlled by the processor (801) according to instructions given by the mobile phone APP.
10. The extraction method of the placental blood is characterized by comprising the following steps:
step one, placing the placenta into a preservation solution of a box body (1), and closing an upper cover (2);
secondly, transporting the sample to a destination laboratory in a constant temperature state;
starting a lifting platform (3) in the biological safety cabinet to lift upwards to press the placenta, and puncturing the placenta by a spurt tissue (4) on the lifting platform (3) to make blood in the placenta flow out;
step four, along with the pressure applied to the placenta by the lifting platform (3), the blood extruded from the placenta and the preservation solution are discharged into a sampling device through a sampling port (7) together, and the extraction of the placenta blood is completed;
the method is realized based on the placenta blood extracting device which integrates transportation and collection as one body and is disclosed by any one of claims 1 to 9.
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