Hair follicle micro organ melanocyte cultivation equipment

Abstract

The invention provides hair follicle micro organ melanocyte cultivation equipment, and relates to the field of cell culture.The hair follicle micro organ melanocyte cultivation equipment comprises a box body, a cultivation box is placed in an inner cavity of the box body, a filtering mechanism is arranged in the inner cavity of the box body and comprises a supporting shell, and a filtering frame is arranged in an inner cavity of the supporting shell in a penetrating mode; a U-shaped plate is connected to the opposite sides of the supporting shells through a rotating shaft, a first vertical pipe is connected to the inner wall of the U-shaped plate through a supporting block, one-way liquid outlet valves are arranged at the bottoms of the two sides of the first vertical pipe in a penetrating mode, one sides of the one-way liquid outlet valves extend to an inner cavity of the filtering frame, and the surfaces of the one-way liquid outlet valves are sleeved with first springs. By arranging a supporting shell, a filtering frame, a U-shaped plate, a first vertical pipe, a one-way liquid outlet valve and a first spring, a nutrient solution enters an inner cavity of the filtering frame, the nutrient solution can be filtered, impurities in the nutrient solution are filtered out, absorption and growth of cells are facilitated, and damage to the cells is avoided.

Description

Technical Field

[0001] The invention relates to the field of cell culture, and in particular to a hair follicle micro-organ melanocyte cultivation device. Background Art

[0002] In the fields of medicine and biology, melanocytes are essential for maintaining normal pigmentation of the skin, hair, and eyes. However, various diseases and conditions, such as vitiligo, mottled disease, and the aging process, may lead to the loss or dysfunction of melanocytes, thereby affecting the individual's appearance and mental health. Traditional treatments, such as medication and phototherapy, although they can relieve symptoms to a certain extent, often cannot fundamentally restore the number and function of melanocytes.

[0003] With the rapid development of stem cell technology and gene editing technology, scientists have begun to explore the use of these advanced technologies to regenerate melanocytes to treat related diseases. Induced pluripotent stem cells (iPSCs) technology allows scientists to reverse somatic cells to a pluripotent stem cell state and then differentiate into various cell types, including melanocytes. These stem cells also have the potential to differentiate into melanocytes. In addition, direct transdifferentiation technology allows scientists to directly convert one somatic cell type into another, such as transdifferentiating fibroblasts into melanocytes, thereby bypassing the stem cell stage. However, how to efficiently generate melanocytes with stable functions and long-term survival capabilities, and how to apply these cells in clinical treatment, are still hot topics and difficulties in current research.

[0004] Therefore, we made improvements to this problem and proposed a hair follicle micro-organ melanocyte cultivation device. Summary of the invention

[0005] The purpose of the present invention is to address the current problem that melanocytes with stable functions and long-term survival ability cannot be efficiently generated and these cells cannot be used in clinical treatment.

[0006] In order to achieve the above-mentioned purpose of the invention, the present invention provides the following hair follicle micro-organ melanocyte cultivation device to improve the above-mentioned problem.

[0007] The specific application is as follows: A hair follicle micro-organ melanocyte cultivation device comprises a box body, a cultivation box is placed in the inner cavity of the box body, a filtering mechanism is arranged in the inner cavity of the box body, the filtering mechanism comprises a supporting shell, a filtering frame is arranged through the inner cavity of the supporting shell, a U-shaped plate is connected to the opposite side of the supporting shell through a rotating shaft, the inner wall of the U-shaped plate is connected to a first vertical pipe through a supporting block, one-way liquid outlet valves are arranged through the bottoms of both sides of the first vertical pipe, one side of the one-way liquid outlet valve extends to the inner cavity of the filter frame, and a first spring is sleeved on the surface of the one-way liquid outlet valve.

[0008] As a preferred technical solution of the present application, the one-way liquid outlet valve is located on one side of the inner cavity of the first vertical tube and is connected to a first inclined plate through a rotating shaft, the top of the first inclined plate is connected to a first connecting ring through a rotating shaft, the bottom of the inner wall of the first vertical tube is connected to a second spring, the top of the second spring passes through the first connecting ring and is connected to a first circular plate, the top of the first vertical tube is sleeved with a second vertical tube, the top of the first circular plate contacts the second circular plate, the top of the second circular plate is connected to a segmented independently controlled electric telescopic rod, the top of the first circular plate passes through and is connected to a telescopic hose, the top of the telescopic hose is connected to a one-way liquid inlet valve, and the surfaces of the first vertical tube and the second vertical tube are connected to a third spring through a support plate.

[0009] As a preferred technical solution of the present application, the top of the second vertical pipe is connected to the first liquid collecting shell, the top of the first liquid collecting shell is movably connected to the second liquid collecting shell, the top of the one-way liquid outlet valve is connected to the bottom of the first liquid collecting shell, the top of the second liquid collecting shell is connected to the first hose, one end of the first hose away from the second liquid collecting shell passes through to the outside of the box body, the right side of the box body is connected to a liquid storage tank, the top of the liquid storage tank is provided with a box cover, one end of the first hose passes through the box cover and extends to the inner cavity of the liquid storage tank.

[0010] As a preferred technical solution of the present application, the top of the support shell is connected to a first guide rod through a support block, the surface of the first guide rod is sleeved with a guide sleeve, the top of the guide sleeve is connected to a second inclined plate through a rotating shaft, the top of the second inclined plate is connected to the first liquid collecting shell through a rotating shaft, the inner cavity of the filter frame is provided with a clamping rod, the inner wall of the support shell is provided with a clamping slot, one end of the clamping rod passes through the filter frame and extends to the inner cavity of the clamping slot, one side of the clamping rod is connected to a fourth spring, one side of the fourth spring is connected to a fixed block, one side of the fixed block is fixedly connected to the inner wall of the filter frame, one side of the clamping rod is connected to a pull rope, and the end of the pull rope away from the clamping rod passes through the fixed block and is connected to a connecting plate.

[0011] The hair follicle micro-organ melanocyte cultivation device prepares hair follicle micro-organ melanocytes through a hair follicle micro-organ melanocyte regeneration method, specifically comprising the following steps: S1. Using induced pluripotent stem cell technology to convert somatic cells into iPSCs and differentiate them into melanocytes; S2, through somatic cell nuclear transplantation, the nucleus of the donor cell is injected into the enucleated oocyte to form a cloned embryo, and then stem cells are isolated from it for the cultivation of melanocytes; S3. Use gene editing tools to modify the genes of melanocytes and use immortalization technology to extend their lifespan; S4, using direct transdifferentiation technology to transdifferentiate a somatic cell type into melanocytes; S5. Use drugs and small molecule compounds to promote the regeneration and functional maintenance of melanocytes; S6. Using bioengineered tissue transplantation methods to construct melanocytes and hair follicle micro-organs into functional tissues and transplant them; In step S3, the CRISPR / Cas9 gene editing tool is used to modify the genes of melanocytes to ensure the continuous secretion of melanin; The step S6 constructs a hair follicle micro-organ with a vascularized structure by 3D bioprinting technology; The induced pluripotent stem cell technology in S1 comprises the following steps: A: Cell source: Skin cells are extracted from human tissues. After subsequent processing, the cells will be reprogrammed into iPSCs; B: Introduction of reprogramming factors: Transcription factors are introduced into somatic cells through viral vector method; C: Induced pluripotency: Factors initiate a series of gene expression changes in cells, causing somatic cells to gradually lose their differentiation characteristics and acquire pluripotency; D: Clone selection and expansion: During the culture process, some cells will gradually form iPSC clones. By selecting, further culturing and expanding these cell clones, iPSC lines can be formed; The induced pluripotent stem cell technology in S1 further comprises the following steps: E: Directed differentiation: using a mixture of biochemical small molecules, growth factors, signaling pathway regulators and cytokines to guide iPSCs to differentiate into melanocytes; F: Result analysis: The differentiated melanocytes were analyzed in terms of morphology, gene expression, and cell function; In step B, the transcription factors include but are not limited to OCT4, SOX2, KLF4 and c-MYC, and the viral vectors include but are not limited to retrovirus, lentivirus and adenovirus.

[0012] Compared with the prior art, the present invention has the following beneficial effects: In the scheme of this application: 1. In order to solve the problem that the existing technology cannot efficiently generate melanocytes with stable functions and long-term survival ability, and cannot apply these cells in clinical treatment, the present application realizes efficient and stable regeneration of melanocytes, and applies them to the reconstruction and transplantation of hair follicle micro-organs. Through induced pluripotent stem cell technology, somatic cell nuclear transfer technology and direct transdifferentiation technology, melanocytes can be efficiently generated from a variety of sources to meet different treatment needs. The genes of melanocytes can be modified by gene editing tools to optimize their functional characteristics, such as enhancing pigmentation ability and improving the ability to resist external damage. At the same time, the application of immortalization technology can prolong the life of melanocytes and reduce the impact of cell aging and apoptosis on the treatment effect; 2. This application can further promote the proliferation and differentiation of melanocytes through the use of drugs and small molecule compounds, while maintaining their normal functions, providing guarantees for long-term survival and effects after transplantation, and constructing regenerated melanocytes and hair follicle micro-organs into functional tissues, and transplanting them, which can restore the pigmentation and hair regeneration ability of damaged parts, thereby significantly improving the appearance and quality of life of patients; 3. By arranging the support shell, the filter frame, the U-shaped plate, the first vertical pipe, the one-way liquid outlet valve and the first spring, the nutrient solution enters the inner cavity of the filter frame, and the nutrient solution can be filtered to filter out impurities in the nutrient solution, which is convenient for cell absorption and growth without causing damage to the cells; 4. By setting a second spring, tension is generated on the first circular plate to facilitate the return of the first circular plate. When the first circular plate moves upward and returns, the pressure below is reduced, so that the one-way liquid inlet valve is opened, and the external nutrient solution enters the bottom of the inner cavity of the first vertical pipe through the one-way liquid inlet valve for replenishment; 5. By setting the fourth spring, tension is generated on the clamping rod, so that the clamping rod extends into the inner cavity of the clamping slot, and then the filter frame is locked and positioned, thereby improving the stability of the filter frame. By cooperating with the fixing block, the pull rope, the connecting plate, the one-way liquid outlet valve, the first inclined plate and the first connecting ring, when the first connecting ring is pressed, the one-way liquid outlet valve pushes the connecting plate to move, and then the clamping rod is driven to move by the pull rope, so that the clamping rod is separated from the clamping slot, which is convenient for disassembly of the filter frame and dumping of impurities in the inner cavity of the filter frame; 6. By arranging a movable plate, a heating ring, a semicircular tube, a special-shaped tube, fan blades, a first one-way air outlet valve, a circular tube and an air outlet net, the heating ring heats the air in the inner cavity of the semicircular tube, and then transports the hot air in the inner cavity of the semicircular tube to the top of the cultivation box to heat the growth environment of the cells. The fan blades rotate to blow the hot air evenly to the cell surface, heating evenly to facilitate cell growth. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 A schematic diagram of the overall structure of the hair follicle micro-organ melanocyte cultivation device provided in this application; Figure 2 A schematic cross-sectional view of the hair follicle micro-organ melanocyte cultivation device provided in this application; Figure 3 A schematic diagram of the filtering mechanism of the hair follicle micro-organ melanocyte cultivation device provided in this application; Figure 4 A cross-sectional schematic diagram of the filtering mechanism of the hair follicle micro-organ melanocyte cultivation device provided in the present application; Figure 5 A schematic diagram of the support shell and filter frame of the hair follicle micro-organ melanocyte cultivation device provided in this application; Figure 6 A schematic diagram of a fourth spring and a fixing block of the hair follicle micro-organ melanocyte cultivation device provided in the present application; Figure 7 A schematic cross-sectional view of the heating mechanism of the hair follicle micro-organ melanocyte cultivation device provided in the present application.

[0014] Indicated in the figure: 1. Box; 2. Cultivation box; 3. Filter mechanism; 30. Support shell; 31. Filter frame; 32. U-shaped plate; 33. First vertical pipe; 34. One-way liquid outlet valve; 35. First spring; 36. First inclined plate; 37. First connecting ring; 38. Second spring; 39. First circular plate; 310. Second circular plate; 311. Segmented independent control type electric telescopic rod; 312. Third spring; 313. Telescopic hose; 314. One-way liquid inlet valve; 315. First liquid collecting shell; 316. Second liquid collecting shell; 317. First hose; 318. Liquid storage tank; 319. First guide rod; 320. Guide sleeve; 321. Second inclined plate; 322. Clamping rod; 323. Clamping groove; 324. Fourth spring; 325. Fixed block; 326. Pull rope; 327. Connecting plate; 328. Support ring; 329. Second vertical pipe; 4. Heating mechanism; 40. Semicircular tube; 41. Special-shaped block; 42. Heating ring; 43. Moving plate; 44. First one-way air inlet valve; 45. Special-shaped tube; 46. First one-way air outlet valve; 47. Second connecting ring; 48. Annular tube; 49. Fifth spring; 410. Sleeve plate; 411. Second guide rod; 412. Telescopic cover; 413. Support frame plate; 414. Partition plate; 415. Horizontal tube; 416. Second one-way air inlet valve; 417. Sixth spring; 418. Second hose; 419. Second one-way air outlet valve; 5. Electric push rod; 6. Support plate. DETAILED DESCRIPTION

[0015] In order to enable those skilled in the art to better understand the scheme of the present invention, the technical scheme in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only 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 should fall within the scope of protection of the present invention.

[0016] As described in the background art, it is impossible to efficiently generate melanocytes with stable functions and long-term survival ability, and it is impossible to apply these cells in clinical treatment.

[0017] In order to solve this technical problem, the present invention provides a hair follicle micro-organ melanocyte cultivation device, which is applied to the field of cell culture.

[0018] Specifically, please refer to Figure 1 , Figure 2 , Figure 3 , Figure 4 , Figure 5 and Figure 6 A hair follicle micro-organ melanocyte cultivation device comprises a box body 1, a cultivation box 2 is placed in the inner cavity of the box body 1, a filtering mechanism 3 is arranged in the inner cavity of the box body 1, the filtering mechanism 3 comprises a supporting shell 30, a filtering frame 31 is arranged through the inner cavity of the supporting shell 30, a U-shaped plate 32 is connected to the opposite side of the supporting shell 30 through a rotating shaft, a first vertical pipe 33 is connected to the inner wall of the U-shaped plate 32 through a supporting block, a one-way liquid outlet valve 34 is arranged through the bottom of both sides of the first vertical pipe 33, one side of the one-way liquid outlet valve 34 extends to the inner cavity of the filtering frame 31, and a first spring 35 is sleeved on the surface of the one-way liquid outlet valve 34.

[0019] By setting the first spring 35, the one-way liquid outlet valve 34 is connected to facilitate the return of the one-way liquid outlet valve 34. By setting the support shell 30, the filter frame 31, the U-shaped plate 32, the first vertical pipe 33, the one-way liquid outlet valve 34 and the first spring 35, the nutrient solution enters the inner cavity of the filter frame 31, and the nutrient solution can be filtered to filter out impurities in the nutrient solution, which is convenient for cell absorption and growth without causing damage to the cells.

[0020] The one-way liquid outlet valve 34 is located on one side of the inner cavity of the first vertical pipe 33 and is connected to the first inclined plate 36 through a rotating shaft. The top of the first inclined plate 36 is connected to the first connecting ring 37 through a rotating shaft. The bottom of the inner wall of the first vertical pipe 33 is connected to the second spring 38. The top of the second spring 38 passes through the first connecting ring 37 and is connected to the first circular plate 39. The top of the first vertical pipe 33 is sleeved with a second vertical pipe 329. The top of the first circular plate 39 contacts the second circular plate 310. The top of the second circular plate 310 is connected to a segmented independently controlled electric telescopic rod 311. The top of the first circular plate 39 passes through and is connected to a telescopic hose 313. The top of the telescopic hose 313 is connected to a one-way liquid inlet valve 314. The surfaces of the first vertical pipe 33 and the second vertical pipe 329 are connected to a third spring 312 through a support plate.

[0021] The top of the segmented independently controlled electric telescopic rod 311 penetrates to the top of the box body 1, and the surface of the first vertical pipe 33 is threadedly connected to the inner wall of the second vertical pipe 329. By setting the second spring 38, tension is generated on the first circular plate 39 to facilitate the return of the first circular plate 39. When the first circular plate 39 moves upward and returns, the pressure below is reduced, so that the one-way liquid inlet valve 314 is opened, and the external nutrient solution enters the bottom of the inner cavity of the first vertical pipe 33 through the one-way liquid inlet valve 314 for replenishment. By setting the telescopic hose 313, the vertical movement of the one-way liquid inlet valve 314 is facilitated. By setting the third spring 312, tension is generated on the first vertical pipe 33 to facilitate the return of the first vertical pipe 33.

[0022] The top of the second vertical pipe 329 is connected to the first liquid collecting shell 315, and the top of the first liquid collecting shell 315 is movably connected to the second liquid collecting shell 316. The top of the one-way liquid outlet valve 34 is connected to the bottom of the first liquid collecting shell 315, and the top of the second liquid collecting shell 316 is connected to the first hose 317. One end of the first hose 317 away from the second liquid collecting shell 316 passes through the outside of the box body 1. The right side of the box body 1 is connected to a liquid storage tank 318, and a box cover is provided on the top of the liquid storage tank 318. One end of the first hose 317 passes through the box cover and extends to the inner cavity of the liquid storage tank 318.

[0023] The first liquid collecting shell 315 is provided to support the second liquid collecting shell 316, and the second liquid collecting shell 316 supports the first hose 317. The first liquid collecting shell 315 and the second liquid collecting shell 316 can rotate relative to each other. The liquid storage tank 318 is provided to store nutrient solution, so as to facilitate timely replenishment of nutrient solution to cells.

[0024] The top of the support shell 30 is connected to the first guide rod 319 through a support block, and a guide sleeve 320 is sleeved on the surface of the first guide rod 319. The top of the guide sleeve 320 is connected to the second inclined plate 321 through a rotating shaft, and the top of the second inclined plate 321 is connected to the first liquid collecting shell 315 through a rotating shaft. The inner cavity of the filter frame 31 is provided with a clamping rod 322, and the inner wall of the support shell 30 is provided with a clamping groove 323. One end of the clamping rod 322 penetrates the filter frame 31 and extends to the inner cavity of the clamping groove 323. One side of the clamping rod 322 is connected to a fourth spring 324, and one side of the fourth spring 324 is connected to a fixed block 325. One side of the fixed block 325 is fixedly connected to the inner wall of the filter frame 31, and one side of the clamping rod 322 is connected to a pull rope 326. The end of the pull rope 326 away from the clamping rod 322 penetrates the fixed block 325 and is connected to a connecting plate 327.

[0025] By setting the first guide rod 319, the guide sleeve 320 is guided to facilitate the lateral movement of the guide sleeve 320. By setting the fourth spring 324, tension is generated on the clamping rod 322, so that the clamping rod 322 extends into the inner cavity of the clamping slot 323, and then the filter frame 31 is locked and positioned, thereby improving the stability of the filter frame 31. By cooperating with the fixing block 325, the pull rope 326, the connecting plate 327, the one-way liquid outlet valve 34, the first inclined plate 36 and the first connecting ring 37, when the first connecting ring 37 is pressed, the one-way liquid outlet valve 34 pushes the connecting plate 327 to move, and then the pull rope 326 drives the clamping rod 322 to move, so that the clamping rod 322 is separated from the clamping slot 323, thereby facilitating the disassembly of the filter frame 31 and the dumping of impurities in the inner cavity of the filter frame 31.

[0026] In order to enable those skilled in the art to better understand the solutions of the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the accompanying drawings.

[0027] It should be noted that, in the absence of conflict, the embodiments of the present invention and the features and technical solutions in the embodiments may be combined with each other.

[0028] It should be noted that similar reference numerals and letters denote similar items in the following drawings, and therefore, once an item is defined in one drawing, further definition and explanation thereof is not required in subsequent drawings.

[0029] Example 1, the hair follicle micro-organ melanocyte cultivation device provided in Example 1 is further optimized, specifically, Figure 7As shown, the top of the second liquid collecting shell 316 is connected to a support ring 328 through a support rod, and the support ring 328 is sleeved on the surface of the segmented independently controlled electric telescopic rod 311. The surface of the segmented independently controlled electric telescopic rod 311 is sleeved with a heating mechanism 4, and the heating mechanism 4 includes a semicircular tube 40, and the inner wall of the semicircular tube 40 is connected to a special-shaped block 41, and the inner wall of the special-shaped block 41 is connected to a heating ring 42. The inner wall of the semicircular tube 40 is threadedly connected to a moving plate 43, and the bottom of the moving plate 43 is connected to There is a first one-way air inlet valve 44, the bottom of the movable plate 43 is connected to a special-shaped tube 45, the bottom of the special-shaped tube 45 is connected to a first one-way air outlet valve 46, the bottom of the first one-way air outlet valve 46 is connected to a second connecting ring 47, the surface of the second connecting ring 47 is sleeved with a circular tube 48, the inner wall of the circular tube 48 is inlaid with an air outlet net, the top of the movable plate 43 is connected to a fifth spring 49, the top of the fifth spring 49 is connected to the top of the inner wall of the box body 1, and the surface of the special-shaped tube 45 is connected to fan blades.

[0030] By setting the fifth spring 49, the movable plate 43 is connected to facilitate the vertical return of the movable plate 43. By setting the air outlet net, the circulation of the gas in the inner cavity of the annular tube 48 with the outside is facilitated. By setting the movable plate 43, the heating ring 42, the semicircular tube 40, the special-shaped tube 45, the fan blades, the first one-way air outlet valve 46, the annular tube 48 and the air outlet net, the heating ring 42 heats the air in the inner cavity of the semicircular tube 40, and then transports the hot air in the inner cavity of the semicircular tube 40 to the top of the cultivation box 2 to heat the growth environment of the cells. The fan blades rotate to blow the hot air evenly to the cell surface, and the heating is even, which is convenient for the growth of the cells.

[0031] Example 2 further optimizes the hair follicle micro-organ melanocyte cultivation device provided in Example 1. Specifically, Figure 7 As shown, both sides of the annular tube 48 are connected with sleeve plates 410, both sides of the inner wall of the box body 1 are connected with second guide rods 411 through support blocks, the sleeve plates 410 are movably sleeved on the surface of the second guide rods 411, the surface of the semicircular tube 40 is connected with a telescopic cover 412, the inner wall of the telescopic cover 412 is connected with a supporting frame plate 413, the end of the telescopic cover 412 away from the semicircular tube 40 is connected with a partition plate 414, the inner cavity of the telescopic cover 412 is provided with a transverse tube 415, and the end of the transverse tube 415 away from the telescopic cover 412 respectively penetrates the telescopic cover 412, the partition plate 414 and the box body 1 and A second one-way air intake valve 416 is connected, and one end of the second one-way air intake valve 416 away from the cross tube 415 is connected to an external oxygen supply device. An air intake hole is provided on the surface of the cross tube 415 and in the inner cavity of the telescopic cover 412. A sixth spring 417 is sleeved on the surface of the cross tube 415, and both sides of the sixth spring 417 are connected to the telescopic cover 412. One end of the telescopic cover 412 away from the semicircular tube 40 is penetrated and connected to a second hose 418. The bottom of the second hose 418 is connected to a second one-way air outlet valve 419, and the bottom of the second one-way air outlet valve 419 is connected to the annular tube 48.

[0032] By setting the sleeve plate 410 and the second guide rod 411, the annular tube 48 is guided to facilitate the vertical movement of the annular tube 48; by setting the support frame plate 413, the telescopic cover 412 is supported so that the telescopic cover 412 can remain in an open state when stretched, so as to facilitate the absorption of external oxygen into the inner cavity of the telescopic cover 412 for replenishment; by setting the second one-way air intake valve 416, the transverse tube 415 and the air intake hole, it is convenient to connect with the external oxygen supply device, so as to facilitate the replenishment of oxygen to the inner cavity of the telescopic cover 412; by setting the sixth spring 417, the semicircular tube 40 is supported to facilitate the return of the semicircular tube 40; by setting the transverse tube 415, the semicircular tube 40 is guided to facilitate the lateral movement of the semicircular tube 40.

[0033] The bottom of the inner wall of the box body 1 is connected to an electric push rod 5, the top of the electric push rod 5 is connected to a support plate 6, the cultivation box 2 is placed on the top of the support plate 6, and the surface of the box body 1 is connected to a box door through a rotating shaft.

[0034] The cultivation box 2 is supported by providing the support plate 6 and the electric push rod 5 , so that the support height of the cultivation box 2 can be adjusted easily.

[0035] The use process of the hair follicle micro-organ melanocyte cultivation device provided by the present invention is as follows: S1. Filtering the nutrient solution: By controlling the operation of the segmented independently controlled electric telescopic rod 311, the segmented independently controlled electric telescopic rod 311 extends at the bottom to drive the second circular plate 310 to move downward, and the second circular plate 310 pushes the first circular plate 39 to move downward, and the pressure below the first circular plate 39 increases, so that the first one-way liquid inlet valve 314 is opened, and the nutrient solution at the bottom of the inner cavity of the first vertical pipe 33 enters the inner cavity of the filter frame 31 through the one-way liquid outlet valve 34, and the nutrient solution is filtered into the filter frame 31. The nutrient solution enters the inner cavity of the cultivation box 2, and the bottom section of the segmented independently controlled electric telescopic rod 311 is retracted to drive the second circular plate 310 to return to its original position, and the second spring 38 pushes the second circular plate 310 to move upward and return to its original position, so that the one-way liquid inlet valve 314 is opened, and the nutrient solution in the inner cavity of the liquid storage tank 318 passes through the first hose 317, the second liquid collecting shell 316, the first liquid collecting shell 315, the one-way liquid inlet valve 314 and the telescopic hose 313 in sequence, and finally enters the bottom of the inner cavity of the first vertical pipe 33 for replenishment, and the cycle repeats; S2, disassembling the filter frame 31: by controlling the extension of the bottom section of the segmented independently controlled electric telescopic rod 311, the second circular plate 310 and the first circular plate 39 are driven to move downward, the first circular plate 39 pushes the first connecting ring 37 to move downward, the first connecting ring 37 pushes the first connecting ring 37 to move horizontally through the first inclined plate 36, the first connecting ring 37 pushes the connecting plate 327 to move, the connecting plate 327 pulls the clamping rod 322 to move through the pull rope 326, so that the clamping rod 322 is separated from the clamping groove 323, the support plate 6 and the cultivation box 2 are driven to move downward by controlling the electric push rod 5 to contract, and the middle section of the segmented independently controlled electric telescopic rod 311 is controlled to extend, so that the first vertical pipe 33, the U-shaped plate 32, and the support shell 30 are driven to move downward, so that the support shell 30 is moved to the bottom of the circular tube 48, and then the filter frame 31 is manually pulled to move and pull it out, and the impurities in the inner cavity of the filter frame 31 are dumped; S3, stirring the nutrient solution: by controlling the contraction of the bottom section of the segmented independently controlled electric telescopic rod 311, the second circular plate 310 is driven to move upward, and the second circular plate 310 drives the first vertical tube 33 and the U-shaped plate 32 to move upward, and the support shell 30 is rotated downward to a vertical state through the support of the second inclined plate 321, and the first vertical tube 33 is connected to the inner wall of the second vertical tube 329 through the threaded connection, and the first vertical tube 33 will also rotate during the upward movement, and the first vertical tube 33 drives the U-shaped plate 32 and the support shell 30 to rotate left and right, and by controlling the extension of the middle section of the segmented independently controlled electric telescopic rod 311, the support shell 30 is driven to extend into the inner cavity of the cultivation box 2, and the nutrient solution is stirred during the rotation of the support shell 30, and the rotation speed of the support shell 30 is controlled to slowly stir the nutrient solution; S4, heating: the temperature of the inner cavity of the box body 1 is detected by the temperature detection module. If the temperature is lower than the preset value, the heating ring 42 is controlled to operate for heating. The middle section of the segmented independently controlled electric telescopic rod 311 is controlled to contract, so as to drive the first liquid collecting shell 315, the second liquid collecting shell 316 and the support ring 328 to move upward. The support ring 328 pushes the movable plate 43 to move upward. The pressure on the movable plate 43 increases, so that the first one-way air outlet valve 46 is opened. The hot air in the inner cavity of the semicircular tube 40 passes through the special-shaped tube 45, the first one-way air outlet valve 46, the annular tube 48 and the air outlet net in sequence and enters the inner cavity of the box body 1. The movable plate 43 rotates while moving upward. The movable plate 43 drives the special-shaped tube 45 and the fan blades to rotate. The fan blades rotate to blow air, and the hot air is evenly blown to the inner cavity of the culture box 2, so that the culture box 2 is evenly heated to ensure the living environment temperature of the cells. S5, oxygenation: the oxygen content in the inner cavity of the box body 1 is detected by the oxygen detection module. When the oxygen content is lower than the preset value, the middle section of the segmented independently controlled electric telescopic rod 311 is controlled to contract, so that the movable plate 43 moves upward, and the movable plate 43 pushes the special-shaped block 41 upward to separate the left and right special-shaped blocks 41, and the special-shaped block 41 drives the semicircular tube 40 to separate, and the semicircular tube 40 moves and compresses the telescopic cover 412 to open the second one-way air outlet valve 419, and the oxygen in the inner cavity of the telescopic cover 412 passes through the second hose 418, the second one-way air outlet valve 419, the annular tube 48 and the air outlet net in turn to increase the oxygen content in the inner cavity of the box body 1, and when the movable plate 43 moves downward, the special-shaped block 41 returns to its original position, pulling the telescopic cover 412 to extend, so that the second one-way air inlet valve 416 is opened, and the oxygen in the inner cavity of the external oxygen supply device passes through the second one-way air inlet valve 416, the cross tube 415 and the air inlet hole in turn, and enters the inner cavity of the telescopic cover 412 for replenishment, and the cycle repeats.

[0036] Embodiment 3 further optimizes the hair follicle micro-organ melanocyte cultivation device provided in Embodiment 1 or 2. Specifically, the hair follicle micro-organ melanocyte cultivation device prepares hair follicle micro-organ melanocytes by a hair follicle micro-organ melanocyte regeneration method, which specifically includes the following steps: S1. Using induced pluripotent stem cell technology to convert somatic cells into iPSCs and differentiate them into melanocytes; S2, through somatic cell nuclear transplantation, the nucleus of the donor cell is injected into the enucleated oocyte to form a cloned embryo, and then stem cells are isolated from it for the cultivation of melanocytes; S3. Use gene editing tools to modify the genes of melanocytes and use immortalization technology to extend their lifespan; S4, using direct transdifferentiation technology to transdifferentiate a somatic cell type into melanocytes; S5. Use drugs and small molecule compounds to promote the regeneration and functional maintenance of melanocytes; S6. Using bioengineered tissue transplantation methods to construct melanocytes and hair follicle micro-organs into functional tissues and transplant them; In step S3, the CRISPR / Cas9 gene editing tool is used to modify the genes of melanocytes to ensure the continuous secretion of melanin; The step S6 constructs a hair follicle micro-organ with a vascularized structure by 3D bioprinting technology; The induced pluripotent stem cell technology in S1 comprises the following steps: A: Cell source: Skin cells are extracted from human tissues. After subsequent processing, the cells will be reprogrammed into iPSCs; B: Introduction of reprogramming factors: Transcription factors are introduced into somatic cells through viral vector method; C: Induced pluripotency: Factors initiate a series of gene expression changes in cells, causing somatic cells to gradually lose their differentiation characteristics and acquire pluripotency; D: Clone selection and expansion: During the culture process, some cells will gradually form iPSC clones. By selecting, further culturing and expanding these cell clones, iPSC lines can be formed; The induced pluripotent stem cell technology in S1 further comprises the following steps: E: Directed differentiation: using a mixture of biochemical small molecules, growth factors, signaling pathway regulators and cytokines to guide iPSCs to differentiate into melanocytes; F: Result analysis: The differentiated melanocytes were analyzed in terms of morphology, gene expression, and cell function; In step B, the transcription factors include but are not limited to OCT4, SOX2, KLF4 and c-MYC, and the viral vectors include but are not limited to retrovirus, lentivirus and adenovirus.

[0037] The present application realizes efficient and stable regeneration of melanocytes, and applies them to the reconstruction and transplantation of hair follicle micro-organs. Through induced pluripotent stem cell technology, somatic cell nuclear transfer technology and direct transdifferentiation technology, melanocytes can be efficiently generated from a variety of sources to meet different treatment needs. The genes of melanocytes are modified by gene editing tools to optimize their functional characteristics, such as enhancing pigmentation ability and improving the ability to resist external damage. At the same time, the application of immortalization technology can prolong the life of melanocytes and reduce the impact of cell aging and apoptosis on the treatment effect.

[0038] In the present invention, unless otherwise clearly specified and limited, the terms "installed", "connected", "connected", "fixed" and the like should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral one; it can be a mechanical connection, an electrical connection, or communication with each other; 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, unless otherwise clearly defined. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0039] Obviously, the embodiments described above are only some embodiments of the present invention, rather than all embodiments. The preferred embodiments of the present invention are given in the accompanying drawings, but they do not limit the patent scope of the present invention. The present invention can be implemented in many different forms. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present invention more thorough and comprehensive. Although the present invention has been described in detail with reference to the aforementioned embodiments, for those skilled in the art, it is still possible to modify the technical solutions recorded in the aforementioned specific implementation methods, or to replace some of the technical features therein with equivalents. Any equivalent structure made using the contents of the specification and drawings of the present invention, directly or indirectly used in other related technical fields, is also within the scope of patent protection of the present invention.

Claims

1. A hair follicle micro-organ melanocyte cultivation device, characterized in that: The invention comprises a box body (1), wherein a cultivation box (2) is placed in the inner cavity of the box body (1), and a filtering mechanism (3) is arranged in the inner cavity of the box body (1), wherein the filtering mechanism (3) comprises a supporting shell (30), wherein a filtering frame (31) is arranged through the inner cavity of the supporting shell (30), wherein a U-shaped plate (32) is connected to the opposite side of the supporting shell (30) via a rotating shaft, wherein the inner wall of the U-shaped plate (32) is connected to a first vertical pipe (33) via a supporting block, wherein one-way liquid outlet valves (34) are arranged through the bottom of both sides of the first vertical pipe (33), wherein one side of the one-way liquid outlet valve (34) extends to the inner cavity of the filtering frame (31), and a first spring (35) is sleeved on the surface of the one-way liquid outlet valve (34).

2. The hair follicle micro-organ melanocyte cultivation device according to claim 1, characterized in that: The one-way liquid outlet valve (34) is located at one side of the inner cavity of the first vertical pipe (33) and is connected to a first inclined plate (36) via a rotating shaft; the top of the first inclined plate (36) is connected to a first connecting ring (37) via a rotating shaft; the bottom of the inner wall of the first vertical pipe (33) is connected to a second spring (38); the top of the second spring (38) passes through the first connecting ring (37) and is connected to a first circular plate (39).

3. The hair follicle micro-organ melanocyte cultivation device according to claim 2, characterized in that: A second vertical tube (329) is sleeved on the top of the first vertical tube (33); the top of the first circular plate (39) contacts the second circular plate (310); the top of the second circular plate (310) is connected to a segmented independently controlled electric telescopic rod (311); a telescopic hose (313) penetrates and is in communication with the top of the first circular plate (39); the top of the telescopic hose (313) is in communication with a one-way liquid inlet valve (314); and the surfaces of the first vertical tube (33) and the second vertical tube (329) are connected to a third spring (312) via a support plate.

4. The hair follicle micro-organ melanocyte cultivation device according to claim 3, characterized in that: The top of the second vertical pipe (329) is connected to the first liquid collecting shell (315), the top of the first liquid collecting shell (315) is movably connected to the second liquid collecting shell (316), and the top of the one-way liquid outlet valve (34) is in communication with the bottom of the first liquid collecting shell (315).

5. The hair follicle micro-organ melanocyte cultivation device according to claim 4, characterized in that: The top of the second liquid collecting shell (316) is connected to a first hose (317); one end of the first hose (317) away from the second liquid collecting shell (316) passes through the outside of the box body (1); the right side of the box body (1) is connected to a liquid storage tank (318); a box cover is provided on the top of the liquid storage tank (318); one end of the first hose (317) passes through the box cover and extends to the inner cavity of the liquid storage tank (318).

6. The hair follicle micro-organ melanocyte cultivation device according to claim 5, characterized in that: The top of the support shell (30) is connected to a first guide rod (319) via a support block; a guide sleeve (320) is sleeved on the surface of the first guide rod (319); the top of the guide sleeve (320) is connected to a second inclined plate (321) via a rotating shaft; and the top of the second inclined plate (321) is connected to the first liquid collecting shell (315) via a rotating shaft.

7. The hair follicle micro-organ melanocyte cultivation device according to claim 6, characterized in that: The inner cavity of the filter frame (31) is provided with a clamping rod (322), the inner wall of the support shell (30) is provided with a clamping groove (323), one end of the clamping rod (322) passes through the filter frame (31) and extends to the inner cavity of the clamping groove (323), and one side of the clamping rod (322) is connected to a fourth spring (324).

8. The hair follicle micro-organ melanocyte cultivation device according to claim 7, characterized in that: One side of the fourth spring (324) is connected to a fixing block (325), one side of the fixing block (325) is fixedly connected to the inner wall of the filter frame (31), one side of the clamping rod (322) is connected to a pull rope (326), one end of the pull rope (326) away from the clamping rod (322) passes through the fixing block (325) and is connected to a connecting plate (327).

9. The hair follicle micro-organ melanocyte cultivation device according to claim 8, characterized in that: The top of the second liquid collecting shell (316) is connected to a support ring (328) via a support rod; the support ring (328) is sleeved on the surface of the segmented independently controlled electric telescopic rod (311); the surface of the segmented independently controlled electric telescopic rod (311) is sleeved with a heating mechanism (4); the heating mechanism (4) comprises a semicircular tube (40); the inner wall of the semicircular tube (40) is connected to a special-shaped block (41); the inner wall of the special-shaped block (41) is connected to a heating ring (42); the inner wall of the semicircular tube (40) is threadedly connected to a moving plate (43); the bottom of the moving plate (43) is connected to the first a one-way air inlet valve (44); the bottom of the movable plate (43) is connected to a special-shaped tube (45); the bottom of the special-shaped tube (45) is connected to a first one-way air outlet valve (46); the bottom of the first one-way air outlet valve (46) is connected to a second connecting ring (47); a circular tube (48) is sleeved on the surface of the second connecting ring (47); an air outlet net is inlaid on the inner wall of the circular tube (48); a fifth spring (49) is connected to the top of the movable plate (43); the top of the fifth spring (49) is connected to the top of the inner wall of the box body (1); and a fan blade is connected to the surface of the special-shaped tube (45).

10. The hair follicle micro-organ melanocyte cultivation device according to claim 9, characterized in that: The hair follicle micro-organ melanocyte cultivation device prepares hair follicle micro-organ melanocytes through a hair follicle micro-organ melanocyte regeneration method, specifically comprising the following steps: S1. Using induced pluripotent stem cell technology to convert somatic cells into iPSCs and differentiate them into melanocytes; S2, through somatic cell nuclear transplantation, the nucleus of the donor cell is injected into the enucleated oocyte to form a cloned embryo, and then stem cells are isolated from it for the cultivation of melanocytes; S3. Use gene editing tools to modify the genes of melanocytes and use immortalization technology to extend their lifespan; S4, using direct transdifferentiation technology to transdifferentiate a somatic cell type into melanocytes; S5. Use drugs and small molecule compounds to promote the regeneration and functional maintenance of melanocytes; S6. Using bioengineered tissue transplantation methods to construct melanocytes and hair follicle micro-organs into functional tissues and transplant them; In step S3, the CRISPR / Cas9 gene editing tool is used to modify the genes of melanocytes to ensure the continuous secretion of melanin; The step S6 constructs a hair follicle micro-organ with a vascularized structure by 3D bioprinting technology; The induced pluripotent stem cell technology in S1 comprises the following steps: A: Cell source: Skin cells are extracted from human tissues. After subsequent processing, the cells will be reprogrammed into iPSCs; B: Introduction of reprogramming factors: Transcription factors are introduced into somatic cells through viral vector method; C: Induced pluripotency: Factors initiate a series of gene expression changes in cells, causing somatic cells to gradually lose their differentiation characteristics and acquire pluripotency; D: Clone selection and expansion: During the culture process, some cells will gradually form iPSC clones. By selecting, further culturing and expanding these cell clones, iPSC lines can be formed; The induced pluripotent stem cell technology in S1 further comprises the following steps: E: Directed differentiation: using a mixture of biochemical small molecules, growth factors, signaling pathway regulators and cytokines to guide iPSCs to differentiate into melanocytes; F: Result analysis: The differentiated melanocytes were analyzed in terms of morphology, gene expression, and cell function; In step B, the transcription factors include but are not limited to OCT4, SOX2, KLF4 and c-MYC, and the viral vectors include but are not limited to retrovirus, lentivirus and adenovirus.

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