Autologous adipose-derived stem cell kit

Abstract

The utility model provides an autologous adipose-derived stem cell kit which comprises a cuboid layered box body, and the layered box body is formed by sequentially connecting a sterile collection layer, an enzymolysis filtering layer, a centrifugal separation layer, a culture amplification layer and a low-temperature cryopreservation layer from top to bottom; the adjacent upper-layer structure and lower-layer structure are separately connected through a sealing buckle, and directional flowing of liquid is achieved through a one-way valve. An injector preloaded with anticoagulant and a blunt-end fat collection needle are arranged in the sterile collection layer, so that tissue damage is reduced. The autologous adipose-derived stem cell kit provided by the utility model can enable the existing kit to have the technical effect of multiple functions, can be used for extracting, amplifying and storing autologous adipose-derived stem cells, and has the technical effects that the whole process is closed, the operation is simple and convenient, and the cell viability can be improved.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically an autologous adipose stem cell kit. Background Technology

[0002] Autologous adipose-derived stem cells (ADSCs) are widely used in tissue repair and anti-aging. They are a type of stem cell with multi-directional differentiation potential isolated from adipose tissue in recent years, possessing advantages such as rapid expansion and resistance to aging common to stem cells. Autologous fat transplantation is an important procedure in cosmetic surgery, widely used in facial plastic surgery, breast augmentation, and other procedures, offering advantages such as no incisions, no rejection reactions, natural appearance, and a natural feel. Traditional extraction methods require multiple steps (such as enzymatic hydrolysis, centrifugation, and medium exchange), which have the following drawbacks:

[0003] (1) The operation is cumbersome and requires professional personnel;

[0004] (2) Open environments are prone to pollution;

[0005] (3) Cell viability is greatly lost during the cryopreservation process.

[0006] Existing reagent kits are mostly limited to a single function (such as only for collection or cryopreservation) and lack a complete integrated design. Therefore, this solution proposes an autologous adipose stem cell kit to solve the above-mentioned technical problems. Utility Model Content

[0007] The purpose of this invention is to provide an autologous adipose stem cell kit, which solves the technical problem of how to make existing kits multifunctional. It can be used for the extraction, expansion and preservation of autologous adipose stem cells, and has the technical effects of being fully closed-loop, easy to operate and improving cell viability.

[0008] An autologous adipose stem cell kit includes a rectangular parallelepiped-shaped layered box. The layered box is composed of a sterile collection layer, an enzymatic hydrolysis and filtration layer, a centrifugation separation layer, a culture and amplification layer, and a cryopreservation layer connected from top to bottom. A cover plate is provided at the top of the sterile collection layer.

[0009] The adjacent upper and lower structures are connected separately by a sealing snap-fit, and the liquid flows in a directional manner through a one-way valve.

[0010] The sterile collection layer contains a pre-filled anticoagulant syringe and a blunt-tipped fat collection needle to reduce tissue damage.

[0011] The enzymatic filtration layer contains collagenase solution and multi-stage filter membranes with pore sizes decreasing from 50μm to 10μm to gradually remove fibrous impurities. The multi-stage filter membranes are arranged in multiple layers that are parallel to each other.

[0012] The centrifugal separation layer integrates detachable centrifuge tubes, an automatic balancing structure, a rotor adapter, and a micro centrifuge. The centrifuge tubes are connected to the rotor adapter, and the two work together to achieve one-button centrifugation.

[0013] The culture and amplification layer includes a pre-filled culture medium and a gas exchange membrane to support cell amplification. The gas exchange membrane is laid flat on the bottom surface of the pre-filled culture medium.

[0014] The cryogenic storage layer contains a programmed cooling cryoprotection tube and a liquid nitrogen storage chamber, with the programmed cooling cryoprotection tube embedded inside the liquid nitrogen storage chamber.

[0015] The programmed cooling cryotube has a built-in temperature sensor and its inner wall is coated with a trehalose film, while the outer side of the tube is wrapped with a phase change material. The programmed cooling cryotube controls the cooling rate through the phase change material to improve the survival rate.

[0016] The multi-stage filter membrane is made of hydrophilic modified polycarbonate material.

[0017] The multi-stage filter membrane comprises three layers with pore sizes of 50μm, 30μm, and 10μm, respectively.

[0018] The beneficial effects of this utility model are as follows:

[0019] (1) This solution is a fully enclosed operation with modular design to reduce the risk of cross-contamination, and the contamination rate is reduced to less than 1%;

[0020] (2) Multi-stage filtration membranes improve cell purity, and programmed cooling cryopreservation tubes ensure cell survival rate ≥90%;

[0021] (3) No special equipment is required, and the operation time is reduced by 50%;

[0022] (4) Applicable to various scenarios such as hospitals and laboratories;

[0023] (5) This utility model discloses an autologous adipose stem cell kit, which adopts a modular layered design and integrates the functions of adipose tissue collection, stem cell separation, expansion and cryopreservation. Through optimized structural design, it solves the problems of cumbersome steps, high risk of contamination and low cell activity of traditional methods. It has the characteristics of simple operation, high efficiency and stability and is suitable for clinical promotion. Attached Figure Description

[0024] Figure 1 This is a schematic diagram of the overall structure of the reagent kit.

[0025] Figure 2 This is a schematic diagram of the internal structure of the enzymatic hydrolysis filter layer.

[0026] Figure 3 This is a schematic diagram of the centrifugal separation layer.

[0027] Figure 4 This is a schematic diagram of the structure of the culture amplification layer.

[0028] Figure 5 This is a schematic diagram of the structure of the cryopreservation layer.

[0029] The attached diagram is labeled as follows: 1. Cover plate; 2. Sterile collection layer; 3. Enzymatic hydrolysis and filtration layer; 31. Multi-stage filter membrane; 4. Centrifugation separation layer; 41. Centrifuge tube; 42. Automatic balancing structure; 5. Culture and amplification layer; 51. Gas exchange membrane; 6. Low temperature cryopreservation layer; 61. Liquid nitrogen storage chamber; 62. Programmed cooling cryopreservation tube. Detailed Implementation

[0030] To more clearly illustrate the technical features of this solution, the following detailed implementation method will be used to explain the solution.

[0031] See Figures 1-5 An autologous adipose stem cell kit includes a rectangular, layered box. The layered box is composed of a sterile collection layer 2, an enzymatic hydrolysis and filtration layer 3, a centrifugation separation layer 4, a culture and amplification layer 5, and a cryopreservation layer 6 connected from top to bottom. A cover plate 1 is provided at the top of the sterile collection layer 2.

[0032] The adjacent upper and lower structures are connected separately by a sealing snap-fit, and the liquid flows in a directional manner through a one-way valve.

[0033] The one-way valve is located at the bottom of the inner layer of each structure and is used for the flow of liquid from top to bottom. For the sake of simplifying the design, it will not be described in detail in the attached drawings.

[0034] The sterile collection layer 2 contains a pre-filled syringe with anticoagulant and a blunt-tipped fat collection needle to reduce tissue damage.

[0035] The enzymatic filtration layer 3 contains collagenase solution and multi-stage filter membrane 31, with the pore size of the filter membrane decreasing from 50μm to 10μm, gradually removing fibrous impurities.

[0036] The centrifugal separation layer 4 integrates a detachable centrifugal tube 41, an automatic balancing structure 42, a rotor adapter, and a micro centrifuge. The centrifugal tube 41 is connected to the rotor adapter, and the two work together to achieve one-button centrifugation.

[0037] In use, the centrifuge tube 41 rotates on the automatic balancing structure 42 and matches the rotor adapter, which is an adapter that connects the centrifuge rotor and the sample tube. Parameters are set and centrifugation is performed; this is the existing technology operation and will not be described in detail here.

[0038] The automatic balancing structure 42 is the balancing disc device, which automatically adjusts the water flow rate by utilizing changes in axial clearance to completely balance the axial force. The balancing disc device consists of a balancing disc, a balancing seat, and an adjusting sleeve. It is existing technology and will not be described in detail here, nor will it be shown in detail in the attached drawings.

[0039] Modern centrifuges typically have an automatic balancing function. In this design, centrifuge tube 41 is connected to automatic balancing structure 42. When in use, automatic balancing structure 42 is positioned on the micro centrifuge.

[0040] The culture and amplification layer 5 includes a pre-filled culture medium and a gas exchange membrane 51. The gas exchange membrane 51 is disposed on the bottom surface of the pre-filled culture medium to support cell amplification.

[0041] The gas exchange membrane 51 is a method and apparatus for improving cell culture conditions in a bioreactor to achieve high-density cell culture. It uses a permeable membrane at the bottom to carry gas exchange to the bottom of the culture medium, thereby improving the transfer efficiency of oxygen and carbon dioxide and supporting high-density culture. This is an existing technology operation and will not be described in detail here.

[0042] The cryogenic storage layer 6 has a built-in programmed cooling cryo-tube 62 and a liquid nitrogen storage chamber 61. The outside of the programmed cooling cryo-tube 62 is covered by the liquid nitrogen storage chamber 61 for cooling.

[0043] The programmed cooling cryotube 62 has a built-in temperature sensor and its inner wall is coated with a trehalose membrane. The outer side of the tube is wrapped with a phase change material. The programmed cooling cryotube 62 controls the cooling rate through the phase change material to improve the survival rate. In use, the low-temperature cryopreservation layer 6 is disassembled from the culture and amplification layer 5, and the programmed cooling cryotube 62 and the liquid nitrogen storage chamber 61 are taken out and placed below the culture and amplification layer 5 to receive the tissue fluid filtered from the gas exchange membrane 51.

[0044] The multi-stage filter membrane 31 is made of hydrophilic modified polycarbonate material.

[0045] The multi-stage filter membrane 31 comprises three filter layers with pore sizes of 50 μm, 30 μm, and 10 μm, respectively.

[0046] The specific working process of this utility model is as follows:

[0047] Adipose tissue is obtained through a collection needle and then injected into the enzymatic filtration layer 3 for digestion.

[0048] After being purified by multi-stage filter membrane 31, the mixture enters centrifugation layer 4, where stem cell precipitate is obtained by centrifugation.

[0049] After the cells were transferred into culture expansion layer 5 for amplification, they were finally cryopreserved in cryopreservation layer 6.

[0050] The technical features of this utility model not described can be implemented by or by using existing technology, and will not be repeated here. Of course, the above description is not a limitation of this utility model, and this utility model is not limited to the examples above. Any changes, modifications, additions or substitutions made by those skilled in the art within the scope of this utility model should also be within the protection scope of this utility model.

Claims

1. An autologous adipose-derived stem cell kit, characterized in that, The container includes a rectangular, layered box, which consists of a sterile collection layer (2), an enzymatic hydrolysis and filtration layer (3), a centrifugation layer (4), a culture and amplification layer (5), and a cryopreservation layer (6) connected from top to bottom. The adjacent upper and lower structures are connected separately by a sealing snap-fit, and the liquid flows in a directional manner through a one-way valve.

2. The autologous adipose-derived stem cell kit according to claim 1, characterized in that, The sterile collection layer (2) contains a syringe pre-loaded with anticoagulant and a blunt-tipped fat collection needle.

3. The autologous adipose-derived stem cell kit according to claim 1, characterized in that, The enzymatic filtration layer (3) contains a collagenase solution and a multi-stage filter membrane (31). The pore size of the filter membrane decreases from 50 μm to 10 μm. The multi-stage filter membrane (31) has multiple layers arranged in parallel to each other.

4. The autologous adipose-derived stem cell kit according to claim 1, characterized in that, The centrifugal separation layer (4) integrates a detachable centrifugal tube (41), an automatic balancing structure (42), a rotor adapter, and a micro centrifuge. The centrifugal tube (41) is connected to the rotor adapter.

5. The autologous adipose-derived stem cell kit according to claim 1, characterized in that, The culture and amplification layer (5) includes a pre-filled culture medium and a gas exchange membrane (51) to support cell amplification. The gas exchange membrane (51) is laid flat on the bottom surface of the pre-filled culture medium.

6. The autologous adipose-derived stem cell kit according to claim 1, characterized in that, The cryogenic storage layer (6) has a built-in programmed cooling cryoprotection tube (62) and a liquid nitrogen storage chamber (61), with the programmed cooling cryoprotection tube (62) embedded inside the liquid nitrogen storage chamber (61).

7. The autologous adipose-derived stem cell kit according to claim 6, characterized in that, The programmed cooling cryopreservation tube (62) has a built-in temperature sensor, its inner wall is coated with a trehalose film and its outer side is wrapped with a phase change material.

8. The autologous adipose-derived stem cell kit according to claim 3, characterized in that, The multi-stage filter membrane (31) is made of hydrophilic modified polycarbonate material.

9. The autologous adipose-derived stem cell kit according to claim 3, characterized in that, The multi-stage filter membrane (31) comprises three layers of filter membrane with pore sizes of 50 μm, 30 μm, and 10 μm, respectively.

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