Rapid extraction kit for mouse bone marrow cells
Through the use of the mouse bone marrow cell rapid extraction kit, the problem that existing devices cannot remove red blood cells and are prone to coagulation is solved, and efficient and rapid bone marrow cell extraction is achieved, which improves cell collection rate and activity.
Patent Information
- Application Number
- CN202421677131.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2024-07-15
- Publication Date
- 2025-05-23
- Estimated Expiration
- 2034-07-15
AI Technical Summary
Existing mouse bone marrow cell extraction devices cannot effectively remove red blood cells in the bone marrow and are prone to coagulation, resulting in a decrease in bone marrow cell collection rate and activity.
A mouse bone marrow cell rapid extraction kit is provided, including a red cleavage tube, a buffer tube, an alcohol tube, a first bone marrow centrifuge tube and a second bone marrow centrifuge tube. Resuspend by alcohol treatment and PBS buffer, remove red blood cells in combination with red blood fluid, and collect bone marrow cells using centrifugal force.
The rapid and effective extraction of mouse bone marrow cells was achieved, the cell collection rate and activity were improved, and red blood cell residues and coagulation problems were avoided.
Smart Images
Figure CN222893169U_ABST
Abstract
Description
Technical Field
[0001] The utility model relates to the field of test kits, in particular to a rapid mouse bone marrow cell extraction test kit. Background Art
[0002] Extracting bone marrow cells from mouse femurs and tibias is a commonly used method in scientific research. The mainstream experimental method is to flush the bone marrow into a culture dish by cutting the epiphysis or fine needle puncture under a sterile environment. The operation process is easy to contaminate bone marrow cells, which is not conducive to subsequent cell culture. The fine needle tip is easy to clog, and the flushing process will introduce residual bone structure, causing bone marrow cell loss, and reducing bone marrow cell production and activity.
[0003] In this regard, some patents currently provide bone marrow cell centrifugal collection devices (CN201721209296.3, CN201320262759.8, CN202022276615.0, CN202022770218.9), which are composed of a small filtering centrifuge tube, a filter device and a large collection centrifuge tube. The working principle is to put the femur and tibia into the upper filtering centrifuge tube, and the bone marrow cells are thrown out from the epiphyseal end by centrifugal force and filtered through the filter once, and then the bone marrow cells are collected by the lower centrifuge tube. These devices can efficiently collect bone marrow cells and avoid cell contamination caused by opening the bone marrow cavity. However, these devices cannot remove the red blood cells in the bone marrow and are prone to coagulation, which causes the subsequent experimental steps to consume some bone marrow cells when removing red blood cells, reducing the bone marrow cell collection rate, and increasing the experimental operation time, which is easy to reduce the activity of bone marrow cells. Utility Model Content
[0004] The utility model is developed to solve the above problems and aims to provide a mouse bone marrow cell rapid extraction kit.
[0005] The utility model provides a mouse bone marrow cell rapid extraction kit, which has the following characteristics and comprises: an erythrocyte lysis tube containing erythrocyte lysis liquid; a buffer tube containing PBS buffer; an alcohol tube containing 75% alcohol; an alcohol plate used for containing alcohol and the femur and tibia of mice; a first bone marrow centrifuge tube used for centrifuging the femur and tibia treated with alcohol, so as to collect bone marrow cells; and a second bone marrow centrifuge tube used for containing erythrocyte lysis liquid and further used for removing red blood cells in bone marrow cells resuspended with PBS buffer.
[0006] The mouse bone marrow cell rapid extraction kit provided by the utility model may also have the following features: a box body, the top of which can be opened, and the red splitting tube, buffer tube, alcohol plate, first bone marrow centrifuge tube and second bone marrow centrifuge tube are all accommodated inside the box body.
[0007] The mouse bone marrow cell rapid extraction kit provided by the utility model may also have such features, including: a protective pad, which is arranged inside the box body and has multiple accommodating holes. The red blood cell splitting tube, the buffer tube, the alcohol tube, the first bone marrow centrifuge tube and the second bone marrow centrifuge tube are respectively inserted into different accommodating holes, so that they can be removably accommodated inside the box body.
[0008] The mouse bone marrow cell rapid extraction kit provided by the utility model may also have the following features, and further include: a buffer dish for containing PBS liquid and the femur and tibia, so as to clean the femur and tibia.
[0009] The mouse bone marrow cell rapid extraction kit provided by the utility model may also have the following features: wherein, the first bone marrow centrifuge tube includes: a first centrifuge tube main body, used to receive and collect bone marrow cells; and a bone holding portion, which is removably arranged on the first centrifuge tube main body and is used to hold the femur and tibia, and an opening is arranged at the bottom thereof for bone marrow cells to pass through.
[0010] The mouse bone marrow cell rapid extraction kit provided by the utility model may also have the following features: wherein, the bone holding portion includes: a first overlapped edge, which is overlapped on the open top of the first centrifuge tube body; and a holding body, which is connected to the first overlapped edge and is arranged inside the first centrifuge tube body, and has an opening at the bottom.
[0011] The mouse bone marrow cell rapid extraction kit provided by the utility model may also have the following features: wherein the containing body has two containing cavities separated from each other, the two containing cavities are used to contain the femur and the tibia respectively, an opening is provided at the bottom of each containing cavity, and a heparin anticoagulant coating is also provided on the bottom inner wall surface of the first centrifuge tube body.
[0012] The mouse bone marrow cell rapid extraction kit provided by the utility model may also have the following features, further comprising: a first filtering part having a first filter screen of 70 um, wherein the first filter screen is connected to the containing body.
[0013] The mouse bone marrow cell rapid extraction kit provided by the utility model may also have the following features: wherein, the second bone marrow centrifuge tube includes: a second centrifuge tube body, used to hold lysed red fluid and bone marrow cells; and a second filter part, which is removably arranged on the second centrifuge tube body and is used to filter the bone marrow cells after being resuspended in PBS buffer.
[0014] The mouse bone marrow cell rapid extraction kit provided by the utility model may also have the following features: wherein, the second bone marrow centrifuge tube includes: a second centrifuge tube body, used to hold lysed red fluid and bone marrow cells; and a second filter part, which is removably arranged on the second centrifuge tube body and is used to filter the bone marrow cells after being resuspended in PBS buffer.
[0015] Functions and effects of utility models
[0016] According to the mouse bone marrow cell rapid extraction kit involved in the utility model, because it includes a red split tube, a buffer tube, an alcohol tube, a red split plate, a first bone marrow centrifuge tube and a second bone marrow centrifuge tube, the alcohol in the alcohol tube can be poured into the alcohol plate first, and then the femur and tibia of the mouse are placed, processed for a short time, and then the femur and tibia are taken out, and the femur and tibia are placed in the first bone marrow centrifuge tube, centrifuged, and bone marrow cells are thrown out from the epiphyseal ends of the femur and tibia by centrifugal force, and then the bone marrow cells are collected at the bottom of the first bone marrow centrifuge tube, the PBS buffer in the buffer tube is poured into the first bone marrow centrifuge tube and the bone marrow cells are resuspended, and then the resuspended liquid is poured into the second bone marrow centrifuge tube, and the second bone marrow centrifuge tube is pre-poured with red split liquid, so that the red blood cells in the resuspended liquid are removed, and bone marrow cells are collected after centrifugation. Therefore, bone marrow cells can be quickly extracted from the femur and tibia of the mouse by the mouse bone marrow cell rapid extraction kit of the utility model, and the collection rate is high and the bone marrow cell activity is strong. BRIEF DESCRIPTION OF THE DRAWINGS
[0017] Figure 1 It is a schematic diagram of the structure of a rapid mouse bone marrow cell extraction kit in an embodiment of the utility model;
[0018] Figure 2 It is a schematic structural diagram of an alcohol tube, a red cracking tube, a buffer tube, an alcohol plate and a buffer plate in an embodiment of the utility model;
[0019] Figure 3 It is a structural schematic diagram of a first bone marrow centrifuge tube in an embodiment of the utility model;
[0020] Figure 4 It is a schematic diagram of the state after the bone containing part of the first bone marrow centrifuge tube in the embodiment of the utility model is taken out;
[0021] Figure 5 It is a schematic structural diagram of a second bone marrow centrifuge tube in an embodiment of the utility model;
[0022] Figure 6 It is a schematic diagram of the state after the second filter part of the second bone marrow centrifuge tube in the embodiment of the utility model is taken out. DETAILED DESCRIPTION
[0023] In the description of this application, it should be noted that, unless otherwise clearly specified and limited, the terms "installed", "connected", and "connected" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection, an electrical connection, or mutual communication; it can be a direct connection, or an indirect connection through an intermediate medium, it can be the internal connection of two elements or the interaction relationship between two elements. For ordinary technicians in this field, the specific meanings of the above terms in this application can be understood according to specific circumstances.
[0024] In order to make the technical means, creative features, objectives and effects of the present invention easy to understand, the following examples are combined with the accompanying drawings to specifically illustrate the mouse bone marrow cell rapid extraction kit of the present invention.
[0025] Example
[0026] Figure 1 It is a schematic diagram of the structure of a rapid mouse bone marrow cell extraction kit in an embodiment of the utility model; Figure 2 It is a schematic structural diagram of an alcohol tube, a red cracking tube, a buffer tube, an alcohol plate and a buffer plate in an embodiment of the utility model; Figure 3 It is a structural schematic diagram of a first bone marrow centrifuge tube in an embodiment of the utility model; Figure 4 It is a schematic diagram of the state after the bone containing part of the first bone marrow centrifuge tube in the embodiment of the utility model is taken out; Figure 5 It is a schematic structural diagram of a second bone marrow centrifuge tube in an embodiment of the utility model; Figure 6 It is a schematic diagram of the state after the second filter part of the second bone marrow centrifuge tube in the embodiment of the utility model is taken out.
[0027] like Figure 1-6 As shown, this embodiment provides a mouse bone marrow cell rapid extraction kit 100, which is used to process the femur and tibia of a mouse to obtain bone marrow cells. The mouse bone marrow cell rapid extraction kit 100 includes a box body 10, a protective pad 20, an alcohol tube 30, a red split tube 40, a buffer tube 50, an alcohol plate 60, a buffer plate 70, a first bone marrow centrifuge tube 80 and a second bone marrow centrifuge tube 90.
[0028] The top of the box body 10 can be opened, and the interior thereof is used to accommodate other components.
[0029] The protective pad 20 is arranged inside the box body 10, and has a plurality of accommodating holes 21. The number of the accommodating holes 21 is 7, and the sizes thereof correspond to the alcohol tube 30, the red splitting tube 40, the buffer tube 50, the alcohol plate 60, the buffer plate 70, the first bone marrow centrifuge tube 80, and the second bone marrow centrifuge tube 90. The alcohol tube 30, the red splitting tube 40, the buffer tube 50, the alcohol plate 60, the buffer plate 70, the first bone marrow centrifuge tube 80, and the second bone marrow centrifuge tube 90 are respectively inserted into the corresponding accommodating holes 21, thereby being arranged on the protective pad 20 and then accommodated in the box body 10. When in use, these components can be taken out of the box body 10.
[0030] like Figure 2 As shown, the alcohol tube 30, the red splitting tube 40 and the buffer tube 50 are tubes with similar structures, and these tubes all have unscrewable caps. The alcohol tube 30, the red splitting tube 40 and the buffer tube 50 contain 75% alcohol (anhydrous ethanol is diluted with distilled water to a concentration of 75%), red splitting solution and PBS buffer respectively. The PBS buffer is weighed to weigh 8.0g NaCl, 0.2g KCl, 1.44g Na 2 HPO 4 , 0.24g KH 2 PO 4 Dissolve in 800mL of distilled water, adjust the solution to 7.4 with HCl, and finally add distilled water to make up to 1L to obtain 0.01M PBS buffer. 2 ). When in use, pour out the corresponding solution in the tube by unscrewing the cap.
[0031] The alcohol dish 60 and the buffer dish 70 are both glass dishes with an open top. The alcohol dish 60 is used to hold the alcohol poured from the alcohol tube 30 and the femur and tibia of the mouse. The buffer dish 70 is used to hold the PBS solution and the femur and tibia of the mouse soaked in alcohol, so as to clean the femur and tibia.
[0032] like Figure 3 As shown, the first bone marrow centrifuge tube 80 is used to centrifuge the femur and tibia after alcohol treatment to collect bone marrow cells. The first bone marrow centrifuge tube 80 includes a first centrifuge tube body 81, a bone containing portion 82 and a first filtering portion 83.
[0033] like Figure 3-4As shown, the first centrifuge tube body 81 is a 1.5 mL transparent plastic centrifuge tube, and the top thereof is provided with an openable first cover 811. The first centrifuge tube body 81 is used to receive and collect bone marrow cells. A sodium heparin anticoagulant coating (heparin dosage 10.0-12.5 IU / mL) is also provided on the bottom inner wall surface of the first centrifuge tube body 81.
[0034] The bone receiving portion 82 is removably disposed on the first centrifuge tube body 81 and is used to receive the femur and tibia. The bone receiving portion 82 includes a first lap 821 and a receiving body 822.
[0035] The first overlapping edge 821 is overlapped on the open top of the first centrifuge tube body 81 .
[0036] The containing body 822 is connected to the first overlap 821 and is arranged inside the first centrifuge tube body 81, and its top is open, and the first cover 811 can also cover the opening. The containing body 822 has two containing chambers separated from each other, and the two containing chambers are used to contain the femur and the tibia respectively. The total capacity of the two containing chambers is 0.5mL. In this embodiment, two containing chambers are formed by providing a longitudinal partition 823 in the containing body 822. The longitudinal partition 823 is arranged in the center of the containing body 822 and is integrally formed with the containing body 822. An opening 824 is provided at the bottom of each containing chamber, and there are two openings in total. The containing body 822 is preferably made of transparent plastic material.
[0037] The first filter part 83 has a first filter screen 831 of 70 um, and the first filter screen 831 is connected to the containing body 822. Specifically, the first filter screen 831 is connected to the containing body 822 through a cylindrical first connecting pipe 832. The first filter screen 831 is a 70 um filter screen.
[0038] The second bone marrow centrifuge tube 90 includes a second centrifuge tube body 91 and a second filter portion 92 .
[0039] The second centrifuge tube body 91 is a 0.5 mL transparent plastic centrifuge tube, and the top of the second centrifuge tube body 91 is provided with an openable second cover 911. The second centrifuge tube body 91 is used to contain lysate and collect bone marrow cells.
[0040] The second filter part 92 is detachably disposed on the second centrifuge tube body 91 and is used to filter the bone marrow cells resuspended in PBS buffer. The second filter part 92 includes a second edge 921 , a second filter screen 922 and a second connecting tube 923 .
[0041] The second overlap 921 is overlapped on the second centrifuge tube body 91. The second filter screen 922 is connected to the first overlap 921 through a cylindrical second connecting tube 923 and is arranged inside the second centrifuge tube body 91. The top of the second connecting tube 923 is open, and the second cover 911 can also cover the opening. The second filter screen 922 is a 40um filter screen.
[0042] The use process of Mouse Bone Marrow Cell Rapid Extraction Kit 100 is as follows:
[0043] Step 1, pour the alcohol in the alcohol tube 30 into the alcohol plate 60, pour the red lysis liquid in the red lysis tube 40 into the second centrifuge tube body 91, take 100uL of the PBS buffer in the buffer tube 50 and pour it into the first centrifuge tube body 81, and take another part and pour it into the buffer plate 70 for use.
[0044] Step 2, remove the mouse femur and tibia from the mouse body and remove the muscles to expose the epiphysis, keep the bone marrow cavity intact, and soak them in 60% alcohol in an alcohol dish for 1-2 minutes.
[0045] Step 3, transfer the femur and tibia soaked in alcohol to PBS buffer in a buffer dish 70, soak for 1 min, and wash off the surface alcohol.
[0046] Step 4, place the bone holding part 82 on the first centrifuge tube body 81, and then the first filter part 83 is also placed in the first centrifuge tube body 81, and then transfer the femur and tibia after washing with PBS to the two holding cavities of the holding body 822 respectively, keeping the epiphyseal end facing down, cover the first cover, use a centrifugal force of 12000g, and instantly separate the bone marrow cells through the 70um first filter 831 and throw them to the bottom of the first centrifuge tube body 81 with a heparin anticoagulant coating below, remove the bone holding part 82 from the first centrifuge tube body 81, and then take out the first filter part 83, and resuspend the bone marrow cells with PBS buffer.
[0047] Step 5, place the second filter part 92 on the second centrifuge tube body 91, add the bone marrow cell suspension into the second filter part 92, let it stand for 1 minute, wait for the bone marrow cell suspension to pass through the 40um second filter 922, then remove the second filter part 92 from the second centrifuge tube body 91, close the second cover 911, turn it upside down to mix, and place it on ice for incubation for 5 minutes, then centrifuge it at 4°C, 450g for 5 minutes, remove the supernatant, wash the cells with PBS buffer, and resuspend the cells to obtain high-quality bone marrow cells for subsequent experiments.
[0048] Functions and Effects of the Embodiments
[0049] According to the mouse bone marrow cell rapid extraction kit involved in the present embodiment, because it includes a red split tube, a buffer tube, an alcohol tube, a red split plate, a first bone marrow centrifuge tube and a second bone marrow centrifuge tube, the alcohol in the alcohol tube can be poured into the alcohol plate first, and then the femur and tibia of the mouse are placed, processed for a short time, and then the femur and tibia are taken out, and the femur and tibia are placed in the first bone marrow centrifuge tube, centrifuged, and bone marrow cells are thrown out from the epiphyseal ends of the femur and tibia by centrifugal force, and then the bone marrow cells are collected at the bottom of the first bone marrow centrifuge tube, the PBS buffer in the buffer tube is poured into the first bone marrow centrifuge tube and the bone marrow cells are resuspended, and then the resuspended liquid is poured into the second bone marrow centrifuge tube, and the second bone marrow centrifuge tube is pre-poured with red split liquid, so that the red blood cells in the resuspended liquid are removed, and bone marrow cells are collected after centrifugation. Therefore, bone marrow cells can be quickly extracted from the femur and tibia of the mouse by the mouse bone marrow cell rapid extraction kit of the utility model, and the collection rate is high and the bone marrow cell activity is strong.
[0050] Furthermore, because the mouse bone marrow cell rapid extraction kit also includes a box body, the top of the box body can be opened, and a protective pad is arranged in the box body, and the protective pad has multiple accommodating holes, the red split tube, the buffer tube, the alcohol tube, the alcohol plate, the buffer plate, the first bone marrow centrifuge tube and the second bone marrow centrifuge tube can be respectively inserted into different accommodating holes, so that they can be removably accommodated in the box body. The above-mentioned various tubes can be stably accommodated in the box body through the protective pad, so that it is easy to use and transport.
[0051] Furthermore, because the mouse bone marrow cell rapid extraction kit has a buffer dish, PBS buffer can be poured into the buffer dish, and then the femur and tibia of the mouse taken out of the alcohol dish are placed in the buffer dish, thereby cleaning the femur and tibia to facilitate the subsequent collection of bone marrow cells.
[0052] Furthermore, because the first bone marrow centrifuge tube includes a first centrifuge tube body and a bone containing portion, the bone containing portion is detachably disposed on the first centrifuge tube body, and the cleaned femur and tibia can be placed in the bone containing portion. During centrifugation, bone marrow cells fall into the bottom of the first centrifuge tube body through the opening, thereby collecting the bone marrow cells.
[0053] Furthermore, because the bottom inner wall surface of the first centrifuge tube body is also provided with a heparin anticoagulant coating, red blood cells can be prevented from coagulation.
[0054] Furthermore, because the bone containing part has a first overlapping edge and a containing body, the entire bone containing part can be conveniently placed on the first centrifuge tube body in a removable manner by overlapping the first overlapping edge on the open top of the first centrifuge tube body.
[0055] Furthermore, because the containing body has two containing cavities separated from each other, the femur and tibia can be placed in the two containing cavities respectively, and the bone marrow cells centrifuged from the femur and tibia respectively fall into the bottom of the first centrifuge tube body through the two openings. The shielding setting can improve the collection rate of bone marrow cells.
[0056] Furthermore, because the mouse bone marrow cell rapid extraction kit also has a first filter part, which has a 70um first filter screen, the first filter screen is connected to the containing body, so as to preliminarily filter the high bone marrow cells falling through the two openings and then fall into the bottom of the first centrifuge tube body.
[0057] Furthermore, because the second bone marrow centrifuge tube includes a second centrifuge tube body and a second filter part, the second filter part has a second overlap and a second filter screen, and the second filter screen is removably arranged on the second centrifuge tube body through the second overlap, the lysed liquid can be poured into the second centrifuge tube body, and then the resuspended high bone marrow cells are poured into the second filter part. After being filtered again by the second filter screen, the bone marrow cells and red blood cells fall into the lysed liquid in the second centrifuge tube body, thereby lysing the red blood cells. After centrifugation, the supernatant is removed to obtain bone marrow cells.
[0058] Those skilled in the art should understand that the present invention is not limited by the above embodiments. The above embodiments and the description are only for explaining the principle of the present invention. Without departing from the spirit and scope of the present invention, the present invention may have various changes and improvements, which are within the scope of the present invention. The scope of protection of the present invention is defined by the attached claims and their equivalents.
Claims
1. A mouse bone marrow cell rapid extraction kit, characterized in that: include: A red splitting tube containing red splitting liquid; a buffer tube containing PBS buffer; Alcohol tube, containing 75% alcohol; An alcohol dish, used for holding the alcohol and the femur and tibia of the mouse; A first bone marrow centrifuge tube, used for centrifuging the femur and tibia after the alcohol treatment to collect bone marrow cells; The second bone marrow centrifuge tube is used to hold lysate and further to remove red blood cells from the bone marrow cells after resuspending in PBS buffer.
2. The mouse bone marrow cell rapid extraction kit according to claim 1, characterized in that: Also includes: The top of the box body can be opened, and the red splitting tube, the buffer tube, the alcohol plate, the first bone marrow centrifuge tube and the second bone marrow centrifuge tube are all accommodated inside the box body.
3. The mouse bone marrow cell rapid extraction kit according to claim 2, characterized in that: Also includes: A protective pad is arranged inside the box body and has a plurality of accommodating holes. The red splitting tube, the buffer tube, the alcohol tube, the first bone marrow centrifuge tube and the second bone marrow centrifuge tube are respectively inserted into different accommodating holes and are thus removably accommodated inside the box body.
4. The mouse bone marrow cell rapid extraction kit according to claim 1, characterized in that: Also includes: A buffer dish is used to contain PBS solution and the femur and the tibia, so as to wash the femur and the tibia.
5. The mouse bone marrow cell rapid extraction kit according to claim 1, Features: Wherein, the first bone marrow centrifuge tube comprises: A first centrifuge tube body, used for receiving and collecting the bone marrow cells; and The bone containing part is detachably disposed on the first centrifuge tube body and is used to contain the femur and the tibia, and an opening is disposed at the bottom thereof for the bone marrow cells to pass through.
6. The mouse bone marrow cell rapid extraction kit according to claim 5, Features: Wherein, the bone holding part comprises: A first lap edge is provided on the top of the opening of the first centrifuge tube body; and The containing body is connected to the first overlapped edge and is arranged inside the first centrifuge tube body, and has the opening at the bottom.
7. The mouse bone marrow cell rapid extraction kit according to claim 6, characterized in that: in, The containing body has two containing chambers separated from each other, and the two containing chambers are used to contain the femur and the tibia respectively. Each of the containing chambers has an opening at the bottom. A heparin anticoagulant coating is also provided on the inner wall surface of the bottom of the first centrifuge tube body.
8. The mouse bone marrow cell rapid extraction kit according to claim 6, characterized in that: Also includes: The first filter part has a first filter screen of 70 um, and the first filter screen is connected to the containing body.
9. The mouse bone marrow cell rapid extraction kit according to claim 8, Features: Wherein, the second bone marrow centrifuge tube comprises: A second centrifuge tube body is used to contain lysate and bone marrow cells; and The second filtering part is detachably arranged on the second centrifuge tube body and is used for filtering the bone marrow cells after being resuspended in PBS buffer.
10. The mouse bone marrow cell rapid extraction kit according to claim 9, Features: Wherein, the second filter unit comprises: A second lap edge is provided on the top of the opening of the second centrifuge tube body; and A second filter screen with a diameter of 40 um is connected to the second overlap and is arranged inside the second centrifuge tube body.