Hair follicle stem cell rapid separation system and separation method

Abstract

The invention belongs to the technical field of cell separation, and particularly relates to a rapid separation system and method for hair follicle stem cells. According to the hair follicle stem cell rapid separation system and method, hair follicle bundles are heated and digested through the arranged dissociation mechanism, so that hair follicles are conveniently separated from hair, the hair follicles are smashed and ground through the arranged grinding mechanism, and hair follicle tissue fragments are obtained; the hair follicle tissue fragments added with the buffer diluent are filtered through the collecting mechanism to obtain the hair follicle cell suspension, and the hair follicle tissue fragments are always at a proper temperature by detecting the temperature of the grinding mechanism and adjusting the rotating speed of the grinding mechanism, so that cell inactivation is avoided.

Description

Technical Field

[0001] The invention belongs to the technical field of cell separation, and in particular relates to a hair follicle stem cell rapid separation system and separation method. Background Art

[0002] Vitiligo is a disease caused by the destruction of melanocytes in the skin, which leads to a lack of melanin in the skin and the formation of local white spots. Its cause may be related to autoimmune diseases, genetics, neurochemicals and other factors. People of different ages, genders and races may suffer from it. The typical symptom is white spots on the skin. When it first occurs, the plaques vary in size and color. If the disease worsens, it can cause white hair.

[0003] In recent years, there are in vitro skin cell rapid separation devices that can obtain skin cells and various active factors in the skin from the healthy parts of the patient's skin, and then apply them to the wound site to accelerate healing. However, the current method of obtaining hair follicle stem cells is generally to separate and culture them through a dissecting microscope or to separate and culture primary cells using a flow cytometer or a magnetic bead sorter. The disadvantage of hair follicle stem cells obtained by these methods is that they cannot be quickly obtained directly in the operating room in a short period of time, and need to be cultured for a long time in the laboratory. The reason is that when hair follicle stem cells are quickly obtained in the operating room, the culture temperature of the hair follicle stem cells cannot be monitored and adjusted, which can easily lead to the inactivation of hair follicle stem cells.

[0004] There is an urgent need to provide a small portable device to solve the problem that the temperature of existing hair follicle stem cells cannot be accurately controlled when processed in the operating room.

[0005] It should be noted that the above information disclosed in this background technology section is only used to understand the background technology of the present application concept, and therefore, the above description is not considered to constitute information of the prior art. Summary of the invention

[0006] The disclosed embodiment at least provides a hair follicle stem cell rapid separation system, including: a separation box, in which a dissociation mechanism, a grinding mechanism and a collection mechanism are respectively arranged;

[0007] The dissociation mechanism is suitable for heating and digesting the hair follicle bundle to separate the hair follicle from the hair;

[0008] The grinding mechanism is suitable for crushing and grinding the separated hair follicles to obtain hair follicle tissue fragments; and

[0009] The collecting mechanism is suitable for filtering the hair follicle tissue fragments after adding the buffer diluent to obtain the hair follicle cell suspension.

[0010] In an optional embodiment, the dissociation mechanism includes: a heating element;

[0011] The heating element is disposed in a dissociation tank provided on the separation box; and

[0012] The heating element is provided with a heat source inside for heating the hair follicle bundles in the digestion and dissociation tank.

[0013] In an optional embodiment, the grinding mechanism includes: a cell grinding component and a control module;

[0014] The cell grinding component is disposed in a grinding groove provided on the separation box; and

[0015] The control module is electrically connected to the cell polishing component to control the polishing speed of the cell polishing component, thereby controlling the temperature of the hair follicle tissue fragments.

[0016] In an optional embodiment, the cell polishing assembly includes: a polishing blade and a temperature measuring guide column;

[0017] The temperature measuring guide post is inserted into the grinding groove, and the grinding blade is inserted into the upper end of the temperature measuring guide post; and

[0018] The bottom of the temperature measuring guide column extends downward from the grinding groove to be connected to the servo motor provided in the separation box; wherein

[0019] The servo motor is suitable for driving the temperature measuring guide column to rotate, and then driving the grinding blade to rotate, so as to grind and break the hair follicle bundle.

[0020] In an optional embodiment, a temperature sensor is provided in the temperature measuring guide column, and the temperature sensor is suitable for obtaining the temperature in the grinding tank; and

[0021] The temperature sensor is electrically connected to the control module to upload the acquired temperature data to the control module;

[0022] The control module is also electrically connected to the servo motor to control the rotation speed of the servo motor according to the acquired temperature data.

[0023] In an optional embodiment, the collection mechanism includes: a cell filter;

[0024] The cell filter is sleeved in the filter slot provided on the separation box; and

[0025] The cell filter is suitable for filtering the hair follicle tissue fragments diluted by buffer, so that the hair follicle cell suspension flows into the filter tank.

[0026] On the other hand, the embodiments of the present disclosure further provide at least one method for rapid separation of hair follicle stem cells, comprising: using the above-mentioned system for rapid separation of hair follicle stem cells; and

[0027] The hair follicle bundle is heated and digested by a dissociation mechanism to obtain the hair follicle;

[0028] The hair follicles are crushed and ground by a grinding mechanism to obtain hair follicle tissue fragments; and

[0029] The hair follicle powder is filtered through a collecting mechanism to obtain a hair follicle cell suspension.

[0030] In an optional embodiment, the method of crushing and grinding hair follicles by a grinding mechanism to obtain hair follicle tissue fragments includes:

[0031] The temperature in the grinding tank is obtained by a temperature sensor arranged in the temperature measuring guide column;

[0032] The rotation speed of the servo motor is controlled according to the temperature in the grinding tank, thereby controlling the rotation speed of the grinding blade.

[0033] In an optional embodiment, the method of controlling the rotation speed of the servo motor according to the temperature in the grinding tank, and thus controlling the rotation speed of the grinding blade includes:

[0034] in

[0035] ω is the real-time speed in RPM, ω0 is the reference speed in RPM, T is the current temperature in °C, T0 is the reference temperature in °C, C is the material coefficient, dimensionless, k is the temperature adjustment coefficient in

[0036] The beneficial effects of the present invention are that the hair follicle stem cell rapid separation system and separation method of the present invention heat and digest the hair follicle bundle through the provided dissociation mechanism, thereby facilitating the separation of the hair follicle from the hair, crushing and grinding the hair follicle through the provided grinding mechanism to obtain hair follicle tissue fragments, and filtering the hair follicle tissue fragments after adding a buffer diluent through the collection mechanism to obtain a hair follicle cell suspension, and through the temperature detection and rotation speed adjustment of the grinding mechanism, the hair follicle tissue fragments are always at a suitable temperature to avoid cell inactivation.

[0037] Other features and advantages of the present invention will be described in the following description, and partly become apparent from the description, or understood by practicing the present invention. The purpose and other advantages of the present invention are realized and obtained by the structures particularly pointed out in the description and the drawings.

[0038] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, preferred embodiments are given below and described in detail with reference to the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0040] Figure 1 A schematic diagram of the three-dimensional structure of a hair follicle stem cell rapid isolation system provided by an embodiment of the present disclosure is shown;

[0041] Figure 2 A schematic diagram of the three-dimensional structure of a grinding mechanism provided by an embodiment of the present disclosure is shown;

[0042] Figure 3 A schematic diagram of the process of the method for rapid isolation of hair follicle stem cells provided in an embodiment of the present disclosure is shown.

[0043] In the figure:

[0044] Separation box 1, grinding mechanism 11, grinding tank 110, temperature measuring guide column 111, grinding blade 112, dissociation mechanism 12, collection mechanism 13. DETAILED DESCRIPTION

[0045] In order to make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution of the present invention will be clearly and completely described below in conjunction with the accompanying drawings. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0046] It should be noted that similar reference numerals and letters represent similar items in the following drawings, so once an item is defined in one drawing, it does not need to be further defined and explained in the subsequent drawings. In addition, in the drawings, the thickness of the components may be exaggerated or reduced in order to effectively describe the technical content.

[0047] Some embodiments of the present invention are described in detail below in conjunction with the accompanying drawings. In the absence of conflict, the following embodiments and features in the embodiments can be combined with each other.

[0048] See also Figure 1 , Figure 1A hair follicle stem cell rapid separation system is shown, comprising: a separation box 1, in which a dissociation mechanism 12, a grinding mechanism 11 and a collection mechanism 13 are respectively arranged; the dissociation mechanism 12 is suitable for heating and digesting the hair follicle bundle to separate the hair follicle from the hair; the grinding mechanism 11 is suitable for breaking and grinding the separated hair follicles to obtain hair follicle tissue fragments; and the collection mechanism 13 is suitable for filtering the hair follicle tissue fragments after adding a buffer diluent to obtain a hair follicle cell suspension.

[0049] In some embodiments, the hair follicle bundle is heated and digested by the dissociation mechanism 12, so as to facilitate the separation of the hair follicle from the hair, the hair follicle is crushed and ground by the grinding mechanism 11, so as to obtain hair follicle tissue fragments, and the hair follicle tissue fragments after adding the buffer diluent are filtered by the collecting mechanism 13 to obtain a hair follicle cell suspension, and by detecting the temperature and adjusting the rotation speed of the grinding mechanism 11, the hair follicle tissue fragments are always at a suitable temperature to avoid cell inactivation.

[0050] In some embodiments, the dissociation mechanism 12 includes: a heating element; the heating element is arranged in a dissociation groove opened on the separation box 1; and a heat source is arranged inside the heating element for heating and digesting the hair follicle bundle in the dissociation groove. Specifically, the heat source includes but is not limited to electric heating. The hair follicle bundle removed by the trephine during the operation is placed in the dissociation groove, and then heated to 37°C for 15 minutes. After the heating is completed, the hair follicle bundle is taken out with ophthalmic forceps and placed on a medical tray. The hair follicles are pulled out along the growth direction of the hair follicles with ophthalmic forceps to obtain the hair follicles.

[0051] See also Figure 2 In some embodiments, the grinding mechanism 11 includes: a cell grinding component and a control module; the cell grinding component is arranged in a grinding groove opened on the separation box 1; and the control module is electrically connected to the cell grinding component to control the grinding speed of the cell grinding component, thereby controlling the temperature of the hair follicle tissue fragments. Specifically, the control module includes but is not limited to using an STM-32 model single-chip microcomputer. Through a temperature sensor preset in the temperature measuring guide column 111, the temperature in the grinding groove 110 can be obtained in real time. Then, the temperature data obtained by the control module is used to adjust the speed of the servo motor, thereby reducing the speed when the temperature is too high to avoid overheating and inactivation of hair follicle cells. However, at the same time, if the speed is too low, sufficient shear force cannot be generated, resulting in the inability to effectively break the cells, thereby affecting the homogeneity of the sample. Therefore, through real-time monitoring of the temperature sensor, the real-time speed of the grinding blade 112 is controlled to reduce the probability of cell inactivation under the premise of sufficient breaking.

[0052] In some embodiments, the temperature measuring guide column 111 is inserted into the grinding groove 110, and the grinding blade 112 is inserted into the upper end of the temperature measuring guide column 111. The bottom of the temperature measuring guide column 111 is connected to the servo motor to drive the grinding blade 112 to rotate to grind and break the hair follicle bundle.

[0053] In some embodiments, the collection mechanism 13 includes: a cell filter; the cell filter is mounted in a filter groove opened on the separation box 1; and the cell filter is suitable for filtering the hair follicle tissue fragments diluted by buffer so that the hair follicle cell suspension flows into the filter groove, and then the hair follicle cell suspension is extracted by a syringe and applied to the wound of the vitiligo patient.

[0054] See also Figure 3 On the other hand, the embodiments of the present disclosure also provide at least one method for rapid separation of hair follicle stem cells, including: using the above-mentioned hair follicle stem cell rapid separation system; and heating and digesting the hair follicle bundles through the dissociation mechanism 12 to obtain hair follicles; crushing and grinding the hair follicles through the grinding mechanism 11 to obtain hair follicle tissue fragments; and filtering the hair follicle powder through the collection mechanism 13 to obtain a hair follicle cell suspension.

[0055] In some embodiments, the method of crushing and grinding hair follicles by the grinding mechanism 11 to obtain hair follicle tissue fragments includes: obtaining the temperature in the grinding groove 110 through a temperature sensor set in the temperature measuring guide column 111; controlling the rotation speed of the servo motor according to the temperature in the grinding groove 110, and then controlling the rotation speed of the grinding blade 112.

[0056] Specifically, the method of controlling the rotation speed of the servo motor according to the temperature in the grinding tank 110 and thus controlling the rotation speed of the grinding blade 112 includes: in

[0057] ω is the real-time speed in RPM, ω0 is the reference speed in RPM, T is the current temperature in °C, T0 is the reference temperature in °C, C is the material coefficient, dimensionless, k is the temperature adjustment coefficient in

[0058] Specifically, since the changes in temperature and speed are not simple linear changes, when the real-time temperature is consistent with the reference temperature, there is no need to adjust the speed. However, when the real-time temperature deviates from the reference temperature, the speed will decrease nonlinearly according to the product of the square of the temperature difference and the material coefficient. When the material coefficient is larger, it proves that the material is more difficult to grind. Therefore, the speed needs to be reduced faster.

[0059] In summary, the hair follicle stem cell rapid separation system and separation method of the present invention heats and digests the hair follicle bundle by the provided dissociation mechanism 12, thereby facilitating the separation of the hair follicle from the hair, crushes and grinds the hair follicle by the provided grinding mechanism 11, thereby obtaining hair follicle tissue fragments, and filters the hair follicle tissue fragments after adding the buffer diluent by the collecting mechanism 13, so as to obtain a hair follicle cell suspension, and by detecting the temperature and adjusting the rotation speed of the grinding mechanism 11, the hair follicle tissue fragments are always at a suitable temperature, thereby avoiding cell inactivation.

[0060] Herein, when it is mentioned that a first component is located on a second component, this may mean that the first component may be directly formed on the second component, or a third component may be interposed between the first component and the second component.

[0061] In this article, when an element or layer is referred to as being "located on," "engaged to," "connected to," "attached to," or "coupled to" another element or layer, it may be directly located on, engaged, connected, attached to, or coupled to another element or layer, or there may be intermediate elements or layers. In contrast, when an element is referred to as being "directly on," "directly engaged to," "directly connected to," "directly attached to," or "directly coupled to" another element or layer, there may be no intermediate elements or layers. Other words used to describe the relationship between elements should be interpreted in a similar manner (e.g., "between" versus "directly between," "adjacent" versus "directly adjacent," etc.). As used herein, the term "and / or" includes any and all combinations of one or more of the associated listed items.

[0062] Herein, example embodiments of the present disclosure will be described in more detail with reference to the accompanying drawings. As used herein, expressions such as "at least one of..." modify the entire list of elements when following a list of elements, rather than modifying individual elements in the list. For example, the expression "at least one of a, b, and c" should be understood to include only a, only b, only c, both a and b, both a and c, both b and c, or all of a, b, and c.

[0063] The terms used herein are only used to describe specific exemplary configurations and are not intended to be limited. As used herein, the singular articles "one", "an" and "the" may also be intended to include plural forms, unless it is clearly indicated above that this is not the case. The terms "comprise", "include" and "have" are inclusive, and therefore specify the presence of features, steps, operations, elements and / or components, but do not exclude the presence or addition of one or more other features, steps, operations, elements, components and / or combinations thereof. The method steps, processes and operations described herein should not be interpreted as necessarily requiring them to be performed in the specific order discussed or shown, unless specifically identified as an execution order. Additional or alternative steps may be adopted.

[0064] As used herein, the phrases "in one embodiment," "according to one embodiment," "in some embodiments," and the like generally refer to the fact that the particular feature, structure, or characteristic following the phrase may be included in at least one embodiment of the present disclosure. Therefore, a particular feature, structure, or characteristic may be included in more than one embodiment of the present disclosure, so that these phrases do not necessarily refer to the same embodiment. As used herein, the terms "example," "exemplary," and the like are used to "serve as an example, instance, or illustration." Any implementation, aspect, or design described herein as "example" or "exemplary" is not necessarily to be construed as preferred or superior to other implementations, aspects, or designs. On the contrary, the use of the terms "example," "exemplary," and the like is intended to present concepts in a concrete manner.

[0065] In the description of the embodiments of the present invention, 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 or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium, or it can be the internal communication of two components. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0066] In the description of the present invention, it should be noted that the orientation or positional relationship indicated by the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inside", "outside", etc. is based on the orientation or positional relationship shown in the drawings, which is only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation of the present invention. In addition, terms such as "first", "second" and other numerical terms do not imply an order or sequence when used in this document unless explicitly indicated above. Therefore, without departing from the teachings of the example embodiments, the first element, component, region, layer or section discussed above may be referred to as a second element, component, region, layer or section.

[0067] Spatially relative terms, such as "inside", "outside", "below", "below", "down", "above", "on", etc., may be used herein to facilitate description of the relationship of one element or feature to another element or feature as illustrated in the figure. In addition to the orientation depicted in the figure, spatially relative terms may be intended to cover different orientations of the device in use or operation. For example, if the device in the figure is turned over, the elements described as "below" or "below" other elements or features will be oriented to be "above" other elements or features. Therefore, the example term "below" can cover the orientation above and below. The device can be oriented in other ways (rotated 90 degrees or in other orientations), and the spatially relative descriptors used herein are interpreted accordingly.

[0068] In the above discussion, unless otherwise stated, the terms "about," "approximately," "substantially," etc., when used to describe a numerical value, mean a variation of + / - 10% of the value.

[0069] Based on the above ideal embodiments of the present invention, the relevant staff can make various changes and modifications without departing from the technical concept of the present invention through the above description. The technical scope of the present invention is not limited to the contents of the specification, and its technical scope must be determined according to the scope of the claims.

Claims

1. A hair follicle stem cell rapid isolation system, characterized in that: include: A separation box, in which a dissociation mechanism, a grinding mechanism and a collection mechanism are respectively arranged; in The dissociation mechanism is suitable for heating and digesting the hair follicle bundle to separate the hair follicle from the hair; The grinding mechanism is suitable for breaking and grinding the separated hair follicles to obtain hair follicle tissue fragments; as well as The collecting mechanism is suitable for filtering the hair follicle tissue fragments after adding the buffer diluent to obtain the hair follicle cell suspension.

2. The hair follicle stem cell rapid isolation system according to claim 1, characterized in that: The dissociation mechanism comprises: a heating member; The heating element is disposed in a dissociation tank provided on the separation box; and The heating element is provided with a heat source inside for heating the hair follicle bundles in the digestion and dissociation tank.

3. The hair follicle stem cell rapid isolation system according to claim 1, characterized in that: The grinding mechanism comprises: a cell grinding component and a control module; The cell grinding component is disposed in a grinding groove provided on the separation box; and The control module is electrically connected to the cell polishing component to control the polishing speed of the cell polishing component, thereby controlling the temperature of the hair follicle tissue fragments.

4. The hair follicle stem cell rapid isolation system according to claim 3, characterized in that: The cell polishing component comprises: a polishing blade and a temperature measuring guide column; The temperature measuring guide post is inserted into the grinding groove, and the grinding blade is inserted into the upper end of the temperature measuring guide post; and The bottom of the temperature measuring guide column extends downward from the grinding groove to be connected to the servo motor provided in the separation box; wherein The servo motor is suitable for driving the temperature measuring guide column to rotate, and then driving the grinding blade to rotate, so as to grind and break the hair follicle bundle.

5. The hair follicle stem cell rapid isolation system according to claim 4, characterized in that: A temperature sensor is arranged in the temperature measuring guide column, and the temperature sensor is suitable for obtaining the temperature in the grinding tank; and The temperature sensor is electrically connected to the control module to upload the acquired temperature data to the control module; The control module is also electrically connected to the servo motor to control the rotation speed of the servo motor according to the acquired temperature data.

6. The hair follicle stem cell rapid isolation system according to claim 1, characterized in that: The collection mechanism comprises: a cell filter; The cell filter is sleeved in the filter slot provided on the separation box; and The cell filter is suitable for filtering the hair follicle tissue fragments diluted by buffer, so that the hair follicle cell suspension flows into the filter tank.

7. A method for rapid isolation of hair follicle stem cells, characterized in that: include: Using the hair follicle stem cell rapid isolation system as described in any one of claims 1 to 6; as well as The hair follicle bundle is heated and digested by a dissociation mechanism to obtain the hair follicle; The hair follicles are crushed and ground by a grinding mechanism to obtain hair follicle tissue fragments; as well as The hair follicle powder is filtered through a collecting mechanism to obtain a hair follicle cell suspension.

8. The method for rapid isolation of hair follicle stem cells according to claim 7, characterized in that: The method of crushing and grinding hair follicles by a grinding mechanism to obtain hair follicle tissue fragments comprises: The temperature in the grinding tank is obtained by a temperature sensor arranged in the temperature measuring guide column; The rotation speed of the servo motor is controlled according to the temperature in the grinding tank, thereby controlling the rotation speed of the grinding blade.

9. The method for rapid isolation of hair follicle stem cells according to claim 8, characterized in that: The method of controlling the rotation speed of the servo motor according to the temperature in the grinding tank, and then controlling the rotation speed of the grinding blade includes: in ω is the real-time speed in RPM, ω0 is the reference speed in RPM, T is the current temperature in °C, T0 is the reference temperature in °C, C is the material coefficient, dimensionless, k is the temperature adjustment coefficient in

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