CN111197025A - Technical method for in vitro culture of melanocyte stem cells for treating leucoderma - Google Patents
Technical method for in vitro culture of melanocyte stem cells for treating leucoderma Download PDFInfo
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- CN111197025A CN111197025A CN202010137384.7A CN202010137384A CN111197025A CN 111197025 A CN111197025 A CN 111197025A CN 202010137384 A CN202010137384 A CN 202010137384A CN 111197025 A CN111197025 A CN 111197025A
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Abstract
The invention discloses a technical method for in vitro culture of melanocyte stem cells in the process of treating leucoderma by hair follicle melanocyte stem cell transplantation, which comprises the following steps: the extraction and separation of hair follicle, the first in vitro culture of melanocyte stem cell, the second in vitro culture of melanocyte stem cell, the culture to enhance the activity of melanocyte, promote the transformation of hair follicle melanocyte stem cell to melanocyte, and the transplantation of melanocyte stem cell. The invention uses the hair follicle in vitro culture technology, puts the extracted and separated individual hair follicle into the special culture solution for culture, and uses the in vitro culture and treatment of the melanocytes of the hair follicle, thereby realizing the shortening of the white spot compound color time and achieving the purpose of quickly curing the leucoderma.
Description
Technical Field
The invention relates to the technical field of hair follicle transplantation, in particular to a technical method for in-vitro culture of hair follicle melanocyte stem cells extracted in the process of treating leucoderma by hair follicle melanocyte stem cell transplantation.
Background
Vitiligo is an acquired skin mucosa pigmentary disorder disease, the color recovery of skin lesion parts is mostly finished by the migration of melanocytes at other parts and the generation of pigments, the skin lesion compound color mode of the vitiligo is summarized into a point type, an edge type, a uniform type and a mixed type, and the vitiligo is not easy to relapse only by realizing the point type compound color. The common clinical vitiligo treatment method mainly comprises traditional Chinese medicine treatment, western medicine treatment such as corticosteroid hormone treatment, photochemotherapy (308 excimer laser) and epidermis transplantation, the former two treatment measures are different from person to person, and the treatment with the traditional Chinese medicine has long treatment course and uncertain curative effect, and some patients have larger side effects after taking the medicine, such as nausea, vomiting, inappetence and the like, and the treatment with the corticosteroid hormone has unstable curative effect and is easy to relapse; the epidermis transplantation, one of the current common methods, is uniform in color, unstable and easy to relapse; 308 excimer laser has long treatment period, general curative effect, great side effect, unstable curative effect even if it is multi-colored, and non-uniform color. In recent years, a single hair follicle transplantation is reported for treating leucoderma, but the therapy has a plurality of defects, so the clinical popularization and application cannot be carried out because: (1) because all the existing hair transplantation treatments for treating the vitiligo are planting at an angle, namely planting hair to the dermis layer of the vitiligo, because the thinking mode of hair planting operation is not jumped out, the transplanted hair follicle is worried not to survive, and the transplanted hair follicle is ignored to be hair follicle melanocyte stem cells; (2) the implanted hair only produces melanin around the hair follicle at the intersection of the white spot and has hair growth, and the hair follicle is cut off below 1/3, so that the process of the delivery and distribution of the melanocyte stem cells of the hair follicle along the outer root sheath cannot be realized, and the process of the white spot color restoration is influenced. The regeneration of pigment around implanted hair follicles can be realized only, the restoration of the color can be realized only by transplanting a large number of hair follicles, the treatment of the vitiligo above a moderate area cannot be completed, and the requirement of the treatment of the vitiligo in a large area cannot be met. The problem of slow color recovery exists although the problem is solved by the transplantation of the hair follicle melanocyte stem cells for treating the leucoderma, and the technical method for in-vitro culture of the melanocyte stem cells for treating the leucoderma effectively solves the problem of slow color recovery after the transplantation of the hair follicle melanocyte stem cells.
Disclosure of Invention
The innovation of the transplantation of the hair follicle melanocyte stem cells for treating the leucoderma is that the transplantation direction is different from that of the transplantation of a single hair follicle for treating the leucoderma. The single hair follicle is transplanted in a manner of being planted in a dermis layer at an angle, and the hair follicle melanocyte stem cell transplantation adopts horizontal direction transplantation; and secondly, the complete external root sheath is reserved, and the melanocyte stem cells of the hair follicle are transplanted to a specific layer of an epidermal layer instead of a dermal layer by taking the hair follicle as a basis. The growth environment and factors of the hair follicle in the epidermis are completely different from those of the dermis, the epidermis is more suitable for melanocyte growth, the dermis is more beneficial to the growth of the hair follicle, and the specific layer of the epidermis is a natural culture medium of melanocyte stem cells of the hair follicle; at present, only 5 hair follicles are needed to be prepared per square centimeter to realize complete color restoration of the area, and the purpose of extracting a small number of hair follicles to realize large-area color restoration is achieved. Specific layers of epidermal layers found in the leucoderma treatment by the hair follicle melanocyte stem cell transplantation can be used as a culture medium of hair follicle melanocyte stem cells, the hair follicle melanocyte stem cells implanted into the prepared hair follicle can be continuously converted into mature melanocytes to secrete melanin, the problem of source of the melanocytes is thoroughly solved, and the white spot polycapsule mouth compound color is realized; however, there is a problem that the time for color restoration is long after transplantation. In view of the above, we invented an in vitro melanocyte culturing method, which can rapidly transform the melanocyte stem cells of hair follicles into mature melanocytes and rapidly produce melanin granules. The invention is the core point of the invention, and the theory and the realization method that the specific layer of the epidermis can be used as the culture medium of the hair follicle melanocyte stem cells and the compound color of the white spot polycystic opening are put forward for the first time in the world. The invention has been verified by electron microscopy.
The dot-shaped hair follicle mouth and hair follicle composite color is realized through innovation of transplanting direction and layers; the problems that the vitiligo with more than the equal area can not be treated and the hair grows after the vitiligo is blackened in the background technology are well solved; the invention achieves the purpose of quickly removing white blood and restoring color by in vitro culture before transplantation of the extracted hair follicle melanocyte stem cells. During the period, more new theories, new theories and new findings of vitiligo treatment are written, and the core part of the people before the papers are published is expected to be protected.
Through clinical observation in recent 3 years and dozens of improvements, the melanin production time is shortened from 2-8 weeks to 1-2 weeks through skin CT (computed tomography) observation on 100 skin lesions, so that the treatment time of the vitiligo is greatly shortened.
The invention aims to provide a technical method for treating leucoderma by hair follicle melanocyte transplantation, which extracts and separates out a single hair follicle by an FUE technology, puts the single hair follicle into a special culture solution for culture, effectively improves the transformation and proliferation of hair follicle melanocyte stem cells to mature melanocytes by culture and ensures the postoperative curative effect; the aim of quickly removing the white spots is achieved through accurate planting, and the problems in the background technology are solved.
In order to achieve the purpose, the invention provides the following technical scheme:
the technical method for culturing the melanocyte stem cells in vitro for treating the vitiligo comprises the following steps:
step S1, extracting hair follicles, extracting a follicular unit, and separating the follicular unit into single hair follicles;
step S2, culturing hair follicles in vitro, placing the extracted and separated individual hair follicles into a special culture solution 1 for culturing, and treating the individual hair follicles by 308 excimer laser to enhance the activity of hair follicle melanocytes and promote the hair follicle melanocytes to differentiate and proliferate to generate black particles, wherein the hair follicle melanocyte stem cells are transformed into mature melanocytes;
step S3, putting the mixture into the culture solution 2 to culture for 1-2 hours, further enhancing the activity of the hair follicle melanocyte stem cells, accelerating the transformation to the melanocytes, promoting mature melanocytes to secrete melanin granules, and culturing for later use;
further, the specific method of step S1 is to extract the hair follicle containing melanocytes of the black hair or beard of the occipital region by using the FUE technique, and separate the hair follicle into individual hair follicles under an electron microscope.
Further, in step S2, the individual hair follicle is extracted and separated by FUE technique, and cultured in special culture solution 1 at 0-6 deg.C for 60 min, and irradiated by excimer laser with wavelength of 308NM 15-20 mj/cm for use.
Furthermore, the main components of the special culture solution are fructus psoraleae injection and normal saline.
Further, in the step S3, a culture solution 2 at 0-4 ℃ is placed for culturing for 1-2 hours, and the main components of the culture solution 2 are dexamethasone sodium phosphate injection and 10% glucose injection.
Compared with the prior art, the invention has the following beneficial effects:
1. the invention extracts and separates out single hair follicle by the hair follicle in-vitro culture technology, extracts the hair follicle containing hair follicle melanocyte stem cells, puts into special culture solution to culture, enhances the activity of the hair follicle melanocyte stem cells, accelerates the transformation to melanocytes, and promotes mature melanocytes to secrete melanin granules. After being cultured and activated, the leucoderma pigmentosum is accurately planted in a leucoderma focus area, so that the source of leucoderma melanocytes is thoroughly solved, the postoperative curative effect is ensured, and the color restoration time is shortened.
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FIG. 1 is provided to provide a further understanding of the present invention and forms a part of the specification, and together with the detailed description of the invention serve to explain the invention and not to limit the invention.
Fig. 1 is a flow chart of the technical method for treating vitiligo by hair follicle melanocyte transplantation, which is used for further explaining the technical method for in vitro culture of melanocyte stem cells in the process of treating vitiligo by hair follicle melanocyte stem cells.
Detailed Description
The following detailed description of the preferred embodiments of the present invention is provided in conjunction with the accompanying drawings of fig. 1, and it is to be understood that the preferred embodiments described herein are merely illustrative and explanatory of the invention and are not restrictive thereof.
The technical key points of the invention are as follows: 1. the in vitro culture technology of hair follicle comprises extracting and separating individual hair follicle by FUE technology, culturing in special culture solution containing dexamethasone injection and normal saline as main ingredients at 0-6 deg.C for 60-180 min, irradiating with excimer laser of wavelength 308NM 50mj, and keeping; the first embodiment is as follows: as shown in fig. 1, the present invention provides a technical solution: the technical method for in vitro culture of the melanocyte stem cells for treating the vitiligo in the process of treating the vitiligo by the autologous hair follicle melanocyte transplantation comprises the following steps: step S2, culturing hair follicles in vitro, placing the extracted and separated individual hair follicles into a special culture solution 1 for culturing, and treating the individual hair follicles by 308 excimer laser to enhance the activity of hair follicle melanocytes and promote the hair follicle melanocytes to differentiate and proliferate to generate black particles, so that hair follicle melanocyte stem cells are transferred to mature melanocytes;
step S3, placing the hair follicle treated in the step S2 into a culture solution 2 to be cultured for 1-2 hours, further enhancing the activity of the hair follicle melanocyte stem cells, accelerating the transformation to melanocytes, promoting mature melanocytes to secrete melanin granules, and culturing for later use;
in the first embodiment:
in this embodiment, in step S2, individual hair follicles are extracted and isolated by FUE technique, and cultured in special culture medium at 0-6 deg.C for 60 min under irradiation of excimer laser with wavelength of 308NM 15-20 mj/cm to increase activity of melanin in hair follicles and convert black cell stem cells into mature melanocytes.
In this embodiment, in step S3, the hair follicle melanocyte stem cells cultured in the special culture medium 1 are placed in the culture medium 2 for culturing for 1-2 hours, so as to further enhance the activity of the hair follicle melanocyte stem cells, accelerate the transformation to melanocytes, promote the mature melanocytes to secrete melanin granules, and culture for later use.
The principle of the invention is as follows: fructus Psoraleae has photosensitivity and can promote melanin transport, excimer laser with wavelength of 308nm can promote melanocyte transformation, and dexamethasone has effects of promoting melanogenesis and regulating immunity. The in vitro matching use can promote the transformation of the melanocyte stem cells to the melanocytes and accelerate the formation of black particles, thereby achieving the purpose of promoting the quick color restoration of white spots.
Although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that changes may be made in the embodiments and/or equivalents thereof without departing from the spirit and scope of the invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (3)
1. A technical method for in vitro culture of melanocyte stem cells in the process of treating leucoderma by hair follicle melanocyte stem cell transplantation,
the method is characterized by comprising the following steps:
step S2, culturing hair follicles in vitro, placing the extracted and separated individual hair follicles into a special culture solution 1 for culturing, and treating the individual hair follicles by 308 excimer laser to enhance the activity of hair follicle melanocytes and promote the hair follicle melanocytes to differentiate and proliferate to generate black particles, so that hair follicle melanocyte stem cells are transferred to mature melanocytes;
step S3, placing the treated hair follicle into the culture solution 2 to be cultured for 1-2 hours, further enhancing the activity of the hair follicle melanocyte stem cells, accelerating the transformation to the melanocytes, promoting mature melanocytes to secrete melanin granules, and culturing for later use;
2. the technical method for in vitro culture of melanocyte stem cells during the treatment of vitiligo of hair follicle melanocyte stem cells according to claim 1, wherein the individual hair follicle extracted and separated in the step S1 is cultured in 0-6 ℃ culture solution 1 for 60 minutes in the step S2, the irradiation of wavelength 308NM excimer laser is 5-20 mj/sq cm for later use, and the main components of the culture solution 1 are Psoralea corylifolia injection and physiological saline; the in-vitro irradiation of the excimer laser with the wavelength of 308NM can accelerate the transformation of the hair follicle melanocyte stem cells to mature melanocytes, and the fructus psoraleae can promote the transportation of the melanocytes.
3. The technical method for in vitro culture of melanocyte stem cells during the treatment of vitiligo according to claim 1, wherein the step S3 comprises culturing the hair follicle treated in step S2 in 0-4 deg.C culture solution 2 for 1-2 hours, wherein the culture solution 2 comprises dexamethasone sodium phosphate injection and 10% glucose injection, and the mixture can promote the transformation of melanocyte stem cells into mature melanocytes and accelerate the proliferation of melanocytes, and the melanocytes are transformed into functional state to produce melanin granules.
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Cited By (2)
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| WO2022151450A1 (en) * | 2021-01-16 | 2022-07-21 | 海口仁术皮肤科门诊部有限公司 | Technical method for treating leucoderma using hair follicle melanocyte stem cell transplantation |
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| CN114522182A (en) * | 2020-11-07 | 2022-05-24 | 海口仁术皮肤科门诊部有限公司 | Preparation method of hair follicle melanin stem cell suspension for treating leucoderma |
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