Application of human iPSC (induced pluripotent stem cell)-sourced platelet-rich suspension in beauty

Abstract

The invention provides a composition for beautifying, which comprises a blood platelet concentrated solution prepared by differentiation of human pluripotent induced stem cells (iPSC), and an additive for preparing a blood platelet suspension or gel, and is characterized in that the composition for beautifying comprises the following components: a blood platelet concentrated solution prepared by differentiation of human pluripotent stem cells (iPSC), and an additive for preparing the blood platelet suspension or gel. The invention aims to provide a functional platelet-rich composition which can be prepared in a large scale, is stable in batch and can be produced in a standardized manner in the aspect of promoting wound healing. According to the composition, the platelet concentrate and platelet-rich plasma (PRP) separated from blood in the composition are completely different in component, the component is stable, and the slow-release growth factor is beneficial to tissue repair.

Description

Technical Field

[0001] The present invention relates to the field of biomedical materials, and in particular to an application of iPSC in vitro prepared platelet-rich materials in cosmetology. Background Art

[0002] The application of platelets in cosmetology is generally achieved in the form of platelet-rich plasma (PRP). The role of PRP is achieved through the interaction and mutual regulation of growth factors. After the growth factors are secreted, they immediately adhere to the surface of the target cell membrane, activate the cell membrane receptors, and stimulate the normal gene sequence expression of the cell. When platelets are activated by thrombin, they can release a large number of growth factors such as TGF-β, PDGF, FGF, HGF and VEGF. PDGF can stimulate the growth of endothelial cells, promote capillary formation, stimulate the chemotaxis of mononuclear macrophages, and increase the ability to synthesize collagen; TGF-β and PDGF can stimulate the exponential proliferation of various cells involved in the repair of the wound, stimulate the synthesis of collagen, and activate the role of macrophages and other cytokines; VEGF is a strong vascular growth factor that plays an important role in wound healing and vascularization, and is a key promoter of angiogenesis in the early stage. In addition to growth factors, PRP also contains fibrin, fibronectin and vitronectin, which form a fiber network that can serve as a scaffold for tissue repair cells, promote cell adhesion and prevent cell loss.

[0003] However, in the field of beauty, the existing technology still lacks functional platelet-rich preparations that can be prepared in large quantities, have stable quality in each batch, and can be produced in a standardized manner.

[0004] The beauty benefits of PRP

[0005] (1) Powerful wrinkle removal. PRP is rich in a variety of growth factors. After being injected into the superficial layer of the dermis, it can stimulate the production of a large amount of collagen, elastic fibers, and colloids, promote the growth and rearrangement of multiple skin tissues, thereby comprehensively improving the skin condition, delaying aging, and achieving the purpose of powerful wrinkle removal. It is commonly used in clinical practice to gradually alleviate and even eliminate wrinkles such as forehead wrinkles, forehead wrinkles, crow's feet, fine lines around the eyes, nasal dorsum wrinkles, nasolabial folds, wrinkles at the corners of the mouth, and neck wrinkles.

[0006] (2) Optimize skin quality. The active factors produced by PRP promote the establishment of skin microcirculation, accelerate metabolism, and comprehensively improve skin quality and complexion, making the skin whiter, more delicate, and more lustrous. It can be clinically used to improve eye bags and dark circles.

[0007] (3) Deep anti-aging. PRP can promote the growth and rearrangement of multiple skin tissues, thereby comprehensively improving the skin condition and continuously delaying aging.

[0008] (4) Lighten age spots. The establishment of facial microcirculation and the acceleration of skin metabolism can prompt the skin to excrete a large amount of toxins on its own, effectively improving various age spots such as pigmentation and dull skin color.

[0009] (5) Lighten age spots. The establishment of facial microcirculation and the acceleration of skin metabolism can prompt the skin to excrete a large amount of toxins on its own, effectively improving various age spots such as pigmentation, solar lentigines, and chloasma.

[0010] (6) Safe anti-allergy. Continuous treatment with PRP will change the original stress system of the skin, making the skin healthier and more energetic, and effectively improving allergic skin. Clinically, it can be used for other allergic diseases such as facial dermatitis.

[0011] (7) Prevent hair loss and promote hair growth. PRP is rich in high-concentration platelets. Its function of promoting cell proliferation, differentiation, and angiogenesis mainly occurs through the degranulation of α-granules after platelets are activated. This process can promote the release of cytokines for hair growth, promote the formation of blood vessels around hair follicles, and increase the blood supply and nutrition of hair follicles. Clinically, it is mostly used for androgenetic alopecia, alopecia areata, etc. Summary of the Invention

[0012] The present invention provides a method for differentiating human pluripotent induced stem cells (iPSC, induced pluripotent stem cell) in vitro, generating platelets using a bioreactor, and activating them in vitro, and finally provides a platelet composition for cosmetic applications.

[0013] To achieve the above object, the present invention provides the following technical solutions:

[0014] The present invention provides a composition for beauty, comprising the following components:

[0015] (1) Platelet concentrate prepared by differentiating pluripotent induced stem cells;

[0016] (2) Additives: normal saline of 0.9% NaCl and 5% human recombinant serum albumin; wherein, the aforementioned composition can be directly injected (into the dermis, superficial subcutaneous layer), smeared, and used in combination of injection and smearing.

[0017] Preferably, the above additives further contain 100 U / mL freeze-dried thrombin, 1% calcium gluconate, and 0.05% type I collagenase.

[0018] The present invention also provides a composition for beauty, comprising the following components:

[0019] (1) Platelet concentrate prepared by differentiating pluripotent induced stem cells;

[0020] (2) Additives: 10 μM ADP, 10 - 100 μg / mL fibrinogen, 2 mM CaCl 2 and 0.05 - 0.5% type I collagenase; among them, the above composition can be directly injected (dermis layer, superficial subcutaneous layer), smeared, and used in combination of injection and smearing.

[0021] Preferably, the concentration of the above platelet concentrate is 5 - 20×10 11 / L.

[0022] Preferably, the above composition is administered once every 1 - 2 months, and every 3 - 4 times is a treatment course.

[0023] More preferably, the above application of the composition in the preparation of drugs for beauty.

[0024] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0025] (1) The process of differentiating iPSCs into hematopoietic precursor stem cells is divided into three stages, and each stage has its corresponding culture medium formula, which helps to improve the purity of CD34 + cells.

[0026] (2) During the process of differentiating hematopoietic precursor stem cells into megakaryocytes, 50 ng / mL IL-21, 5 nM Tazemetostat, 200 nM

[0027] Eltrombopag and 200 nM iBET151 are added to the culture medium, which can promote the differentiation and proliferation of megakaryocytes.

[0028] (3) After the megakaryocytes are cryopreserved and thawed, and before using the reactor, add

[0029] 0.5 ng / mL collagen and 5 nM Fingolimod HCl, which helps to promote platelet maturation.

[0030] (4) The reactor for generating platelets has a high-density grid structure, and the function of the grid structure is to cut the liquid flow, which can produce intense megakaryocyte fragmentation and help release platelets.

[0031] (5) Compared with PRP separated from blood, the iPSC platelet preparation is relatively stable in composition, and the slow-release growth factor helps tissue repair.

[0032] (6) The preparation methods (suspension formula, preparation formula of activated suspension, gel preparation formula and preparation method) have never been used in the application scenario of iPSC-platelets. Description of the Drawings

[0033] The accompanying drawings forming a part of this application are used to provide a further understanding of the present invention. The schematic embodiments and descriptions thereof of the present invention are used to explain the present invention and do not unduly limit the present invention. In the drawings:

[0034] Figure 1 are the morphology (a) of hematopoietic progenitor cells generated from pluripotent induced stem cells (iPSCs), the cell size (b) of hematopoietic stem cells, and CD34 obtained by passage purification + cells (c) and their purity (d). CD34: A marker mainly expressed on hematopoietic stem cells and hematopoietic progenitor cells. BF: brightfield, bright field view.

[0035] Figure 2 are the comparisons of various data of induced megakaryocytes (iMKs) derived from iPSCs of a newborn male (A) and a 34-year-old female (B) after cryopreservation and resuscitation, including morphology (b, c), proliferation ability (d, e), cell viability (f), and cell size (g).

[0036] Figure 3 are the in vitro maturation and shedding process of platelets (a), and the proportion (b), overall size (c) of platelets produced by a single megakaryocyte in static and reactor environments; through morphological observation, it is found that intense megakaryocyte fragmentation and platelet release can occur in the reactor within 72 hours (d). Through flow cytometry detection, it is found that iPSC-derived platelets (iPLTs) have a similar marker expression (f) compared with peripherally blood-derived platelets (e). BF: bright field, bright field view. PB-PLTS: Peripherally blood-derived platelets.

[0037] Figure 4 are the states of non-activated and activated platelet suspensions during the preparation of platelet suspensions. Detailed implementation manners

[0038] The following further describes the present invention in combination with the accompanying drawings and specific embodiments. The protection scope of the present invention is not limited to the following embodiments. It should also be understood that the terms used in the embodiments of the present invention are for the purpose of describing specific specific implementation manners, rather than for limiting the protection scope of the present invention, and are not uniquely defined. Without departing from the spirit and scope of the inventive concept, changes and advantages that can be conceived by those skilled in the art are included in the present invention, and the appended claims and any equivalents thereof are the protection scope of the present invention.

[0039] All technical and scientific terms used in this document have the same meaning as commonly understood by those skilled in the art to which this invention pertains. In other cases, certain terms used herein will have their meanings clarified in the specification. For experimental methods without specific conditions noted in the following examples, they are all common knowledge and well-known general knowledge to those skilled in the art. The examples in this application and the features in the examples can be combined with each other.

[0040] The features and advantages of the present invention can be further understood through the following detailed description in conjunction with the accompanying drawings. The provided examples are only illustrative of the methods of the present invention and do not limit the remaining content disclosed by the present invention in any way.

[0041] In order to provide a platelet preparation with stable components, as well as a preparation method that can be mass-produced, batch-stable, and standardized, the present invention utilizes the iPSC platelet preparation method, reactor, suspension formulation, activated suspension formulation, gel formulation, and preparation method to achieve technical effects.

[0042] The present invention provides a method for preparing platelets using stem cells. First, the process of differentiating induced pluripotent stem cells (iPSC) into hematopoietic progenitor stem cells (HPC) is divided into three stages, and each stage has its corresponding culture medium formulation:

[0043] Table 1.

[0044]

[0045]

[0046] Secondly, during the process of differentiating HPC into megakaryocytes (iMK), adding 50 ng / mL IL-21, 5 nM Tazemetostat, 200 nM Eltrombopag, and 200 nM iBET151 to the culture medium can promote the differentiation and proliferation of megakaryocytes.

[0047] Finally, after cryopreservation and resuscitation of megakaryocytes and before using the reactor, add a maturation process from static to dynamic (see points 1 and 2 in Example 3), and add 0.5 ng / mL Collagen and 5 nM Fingolimod HCl to promote platelet maturation.

[0048] The present invention uses a reactor to generate platelets. Among them, the reactor has a high-density grid structure, and the function of the grid structure is to cut the liquid flow. There are two ways for the grid structure to cut the liquid flow: 1) the grid is static and the liquid flow is dynamic; or 2) the grid is dynamic and the liquid flow is static.

[0049] Although the method for preparing iPSC platelet preparations in the present invention refers to the traditional PRP preparation concept, the iPSC platelet preparations and PRP separated from blood are completely different in composition, and the composition of PRP separated from blood is relatively unstable. Traditional iPSC platelets include a large proportion of plasma components. Due to different individual / blood bank sources, there are significant differences in their composition and proportion, which can lead to differences in the effects during application; the composition of iPSC-platelets is determined, the batches are stable, there are no compositional differences caused by different sampling individuals, and they can be prepared on a large scale and strictly quality controlled, so as to obtain a standardized product with strict control over the production process and product quality. Therefore, it can play a very good guiding and reference effect on the dosage and effect of clinical application. Correspondingly, for example: The platelet-rich suspension product derived from iPSC is different from peripheral blood PRP, and it does not contain white blood cell components and plasma components. The platelet-rich suspension derived from iPSC contains iPSC-induced platelets and 5% albumin components. The platelet-derived growth factors produced in large quantities after activation can play a role in repairing tissue damage. Especially for the gel-like PRP, due to the fibrous interweaving and winding of components such as platelets and growth factors, it can play a sustained-release effect, so as to achieve a relatively long-term use effect and also has the effect of tissue repair.

[0050] The suspension and gel additives of the present invention are formulated as follows:

[0051] Table 2.

[0052]

[0053] In addition, the iPSC platelet preparations of the present invention can be applied to different scenarios, such as treating arthritis, tennis elbow, promoting wound healing, treating alopecia, etc.; they can also be applied to the beauty field.

[0054] Example 1: Differentiate iPSC into hematopoietic progenitor cells (iHPCs)

[0055] The iPSC of the present invention is sampled from hospital skin biopsies, and the primary fibroblasts are cultured in the laboratory and screened to obtain an ideal iPSC cell line after reprogramming.

[0056] 1. Dissociate iPSC into single cells and evenly seed them in a culture dish pre-coated with human recombinant laminin (rhLaminin-521, Thermofisher). The culture medium is Essential 8 (Thermofisher) supplemented with 30 ng / mL VEGF-A165, 10 ng / mL bFGF, 5 μM Y-27632, 2 mM Chir-99021, and 20 ng / mL Activin A, and the culture time is 2 days.

[0057] 2. Then change the culture medium to HDM, and the formula is as follows:

[0058] HDM culture medium: Add 1x ITS, 1x glutamax, 0.45 mM monothioglycerol, 50 μg / mL ascorbic acid, and 20% KO-SRM to Iscove modified Dulbecco (IMDM).

[0059] After that, add 30 ng / mL VEGF-A165, 10 ng / mL bFGF, 20 ng / mL BMP4, 5 μM Y-27632, and 20 ng / mL Activin A, and continue culturing for 4 days.

[0060] 3. Then change the culture medium to HDM culture medium, and add 30 ng / mL VEGF-A165, 10 ng / mL bFGF, 50 ng / mL stem cell factor (SCF), 50 ng / mL thrombopoietin (TPO), 2 U / mL erythropoietin (EPO), and 5 μM Y-27632. Optionally, further add 50 ng / mL IL-3 and 50 ng / mL IL-6, and continue culturing for 8 days.

[0061] 4. Collect the hematopoietic progenitor stem cells (iHPCs) growing in suspension, collect the culture medium supernatant containing iHPCs, centrifuge at 1000 rpm for 5 minutes.

[0062] The results are shown in Figure 1 . The hematopoietic progenitor stem cells (Hematopoietic progenitor cells, HPC) generated from iPSCs in this example Figure 1 a) have a cell size similar to that of CD34 cells isolated from peripheral blood + cells ( Figure 1 b), and through passage purification, the obtained CD34 + cells ( Figure 1 c) can reach a purity of about 90% ( Figure 1 d). More starting materials help reduce interference in the subsequent differentiation process and do not produce excessive miscellaneous cells.

[0063] Example 2: Differentiation and proliferation of megakaryocytes

[0064] I. Differentiate hematopoietic progenitor stem cells (iHPC) into megakaryocytes (iMKs)

[0065] 1. Collect the hematopoietic precursor stem cells obtained in Example 1 and seed them in a culture dish pre-coated with 0.1% gelatin. The culture medium is HDM medium supplemented with 20 ng / mL VEGF-A165, 5 ng / mL bFGF, 50 ng / mL thrombopoietin (TPO), 20 ng / mL IL-3, 20 ng / mL IL-6, 50 ng / mL IL-21, 5 nM Tazemetostat, 200 nM Eltrombopag, and 200 nM iBET151, and continue culturing for 15 - 20 days.

[0066] 2. Collect the suspended megakaryocytes.

[0067] II. In vitro proliferation of megakaryocytes (iMKs)

[0068] 1. Transfer the megakaryocytes obtained in the previous step I. to a low-attachment culture flask / shaker flask, change the culture medium to HDM supplemented with 50 ng / mL thrombopoietin (TPO), 50 ng / mL stem cell factor (SCF), 50 ng / mL IL-21, and 5 μM Y-27632, and place it on a horizontal shaking incubator for amplification, with the rotation speed adjusted to 120 - 150 rpm.

[0069] 2. Megakaryocytes can be amplified in vitro for several months while maintaining their properties unchanged. The passage cycle is 3 days, and the seeding density each time is 5×10 6 / mL.

[0070] 3. Megakaryocytes can be cryopreserved. The cryopreservation solution is HDM + 5% BSA + 5% DMSO, and after programmed freezing, it is stored in liquid nitrogen.

[0071] For the results, please see Figure 2 . Two different background iPSCs ( Figure 2 a) megakaryocytes (iMKs) derived from a newborn male (A) and a 34-year-old female (B), after cryopreservation and resuscitation, can maintain a dispersed and uniform morphology during long-term dynamic proliferation culture ( Figure 2 b, Figure 2 c), a proliferation ability of 10,000-fold in 15 days ( Figure 2 d, Figure 2 e), as well as relatively stable cell viability ( Figure 2 f) and cell size ( Figure 2 g).

[0072] Example 3: Production of platelets

[0073] 1. Suspend the megakaryocytes obtained in Example 2 in PM medium, and add 50 ng / mL thrombopoietin (TPO) and 5 μM Y-27632, and statically culture for 1 day.

[0074] PM medium: Add 1x ITS, 1x glutamax, 0.45 mM monothioglycerol, 50 μg / mL ascorbic acid, 10 U heparin, and 5% human plasma to Iscove modified Dulbecco (IMDM).

[0075] 2. Replace the medium with PM medium, add 50 ng / mL thrombopoietin (TPO), 0.5 ng / mL collagen, 5 nM Fingolimod HCl, and 5 μM Y-27632, incubate statically for 1 day, and then continue to culture on a horizontal shaking incubator for 1 day with the rotation speed adjusted to 120 - 150 rpm.

[0076] 3. Transfer the megakaryocytes obtained in the above steps to a bioreactor for platelet maturation in a 1 L system, and continue to culture for 5 - 6 days at 37°C and 5% CO 2 , and the maximum horizontal liquid streamline velocity in the reactor is 30 cm / s.

[0077] The specifications of the reactor are as follows: The appearance is a cylindrical structure, the height of the external tank is 160 mm, the bottom diameter is 130 mm, and the thickness of the culture container is 5 mm; it is embedded with a multi-layer movable grid structure, with a variable-direction rotational movement mode, a rotational speed of 120 - 150 rpm, and a maximum liquid flow linear velocity of 30 cm / s.

[0078] For the results, please refer to Figure 3 . The in vitro maturation and shedding process of platelets in this example( Figure 3 a), in the parallel control of the static environment and the reactor environment, the proportion of platelets produced by a single megakaryocyte under reactor conditions can reach more than 1:100( Figure 3 b), and the overall size is closer to platelets from peripheral blood( Figure 3 c). Through morphological observation, it was found that intense megakaryocyte fragmentation and platelet release can occur within 72 hours in the reactor( Figure 3 d). Through flow cytometry detection, it was found that iPSC-derived platelets (iPLTs) and platelets from peripheral blood( Figure 3 e) have similar marker expression( Figure 3 f) conditions.

[0079] Example 4: Preparation method of platelet suspension and gel

[0080] 1. Platelet-rich suspension derived from human iPSCs

[0081] After concentrating and purifying the iPSC platelets in Example 3, they were suspended in physiological saline containing 0.9% NaCl at a concentration of 5 - 20×10 11 / L, and 5% human recombinant serum albumin (rhHSA) was added thereto.

[0082] 2. Activated human iPSC-derived platelet-rich suspension

[0083] Based on the suspension in the above point 1, 100 U / mL freeze-dried thrombin, 1% calcium gluconate and 0.05% type I collagenase were added.

[0084] 3. Human iPSC-derived platelet-rich gel

[0085] The iPSC platelets obtained in Example 3 were concentrated to 1×10 11 / L or more, suspended in PBS, and 10 μM ADP, 10 - 100 μg / mL fibrinogen, 2 mM CaCl 2 and 0.05 - 0.5% type I collagenase were added to the solution. It was placed in a sterile environment at room temperature and allowed to stand and crosslink for 5 minutes. Centrifuged at 200 rpm for 15 minutes, and then continued to centrifuge at 1500 rpm for 2 minutes. The supernatant was discarded to obtain the lower layer rich in activated platelet gel.

[0086] For the results, please refer to Figure 4 . During the preparation of the platelet suspension, the unactivated platelet suspension (described in method 1 of this example) was in a relatively clear state. After activation (described in method 2 of this example), due to the aggregation of platelets and the entanglement of fibrin, the suspension immediately became turbid (the control group was platelets isolated from peripheral blood). Among them, compared with the platelets isolated from peripheral blood in the control group, the aggregation effect of iPSC platelets was better.

[0087] Example 5: Application of platelet suspension in the treatment of arthritis

[0088] 1. The preparation method of the human iPSC-derived platelet-rich suspension was as in Examples 1 - 4.

[0089] 2. The above platelet suspension was used to treat arthritis, specifically including osteoarthritis and rheumatoid arthritis.

[0090] 3. The injection method was: intra-articular injection into the knee joint under ultrasound guidance, using a 22-gauge needle, 5 mL of fresh human iPSC-derived platelet-rich suspension each time. After injection, the knee joint was passively flexed and extended 5 times, and the patient rested for 10 minutes. Injected once a week, 3 times per course of treatment. The patients were followed up within 12 months after injection.

[0091] 4. The evaluation methods for treatment improvement are as follows: 1) average knee joint pain score (11-point numerical rating scale, 0 = no pain - 10 = most severe pain); 2) measurement of the medial tibial cartilage volume by MRI; 3) 31 auxiliary evaluation indicators (25 related to symptoms and 6 for MRI evaluation) to evaluate pain, function, quality of life, overall changes, and joint structure, etc.

[0092] Example 6: Application of platelet suspension in the treatment of tennis elbow

[0093] 1. The preparation method of human iPSC-derived platelet-rich suspension is as in Examples 1 to 4.

[0094] 2. The above platelet suspension is used to treat tennis elbow.

[0095] 3. The treatment method is as follows: The injection site is blocked with 0.5% bupivacaine and adrenaline, and then 2 - 3 mL of the prepared human iPSC-derived platelet-rich suspension is injected into the extensor carpi radialis brevis tendon and the surrounding area by the tendon puncture method. The needle is inserted once without being withdrawn and the injection direction is changed 5 times to infiltrate the tendon.

[0096] 4. The evaluation methods for treatment improvement are as follows: 1) At 4, 8, 12, 16, 20, and 24 weeks after treatment, if the pain score improvement of the VAS pain score for wrist extension against resistance (judging the severity of pain by the visual analogue method, VASRWE) is 25% or more compared with the baseline, it is considered a success; 2) Auxiliary evaluation indicators: Patient-rated Tennis Elbow Evaluation Questionnaire (PRTEE); Extended wrist examination.

[0097] Example 7: Application of platelet suspension in the treatment of wound healing

[0098] 1. The preparation method of human iPSC-derived platelet-rich suspension is as in Examples 1 to 4.

[0099] 2. The above suspension is used to promote wound healing, such as in the healing of trauma and burns.

[0100] 3. The treatment method is as follows: After surgical treatment of the trauma, before closing and suturing the incision, the iPSC-derived platelet-rich suspension is applied into the wound, with at least 5 mL at each site and the platelet content not less than 1×10 6 / μL, and the dosage is adjusted according to the trauma area and the degree of injury.

[0101] 4. The evaluation indicators for treatment improvement are as follows: 1) Wound healing time; 2) Wound infection assessment; 3) Wound healing quality.

[0102] For general wounds, unactivated platelet PRP is used to enable the slow release of growth factors to promote healing. For severe injuries or wounds with an infection tendency, activated platelets are used to enrich a large amount of growth factors at the wound site and release them rapidly, thus quickly promoting wound healing.

[0103] Example 8: Application of platelet suspension in the cosmetic field

[0104] 1. The preparation method of human iPSC-derived platelet-rich suspension is as described in Examples 1 to 4.

[0105] 2. The above suspension is used in the cosmetic field.

[0106] 3. The treatment methods are: direct injection (dermis layer, superficial subcutaneous layer), application, and combined use. According to different application directions, it can be referred to that the treatment is carried out once every 1 to 2 months, and 3 to 4 times for each course of treatment.

[0107] 4. Precautions for treatment:

[0108] The treatment with human iPSC-derived platelet-rich suspension does not require a recovery period, and normal work can be resumed the day after treatment. After treatment, swelling or bleeding points may appear in the treatment area. Generally, mild swelling can subside on the same day, and bleeding points can generally disappear in 2 to 3 days. It should be noted that:

[0109] (1) Women should prepare and use concentrated platelet products during non-menstrual periods. Pregnant, lactating, and women planning pregnancy, as well as patients with anemia, abnormal blood coagulation function, and abnormal liver function, are prohibited from using;

[0110] (2) Keep the treatment area dry for 24 hours after treatment, do not get it wet, do not use other irritating skin care products; pay attention to sun protection, do not take anticoagulant drugs 2 weeks before treatment; it is not recommended to take a steam bath or massage 1 week after treatment.

[0111] Example 9: Application of platelet suspension in the treatment of hair loss

[0112] 1. The preparation method of human iPSC-derived platelet-rich suspension is as described in Examples 1 to 4.

[0113] 2. The above suspension is used to treat hair loss.

[0114] 3. The treatment methods are: direct multi-point injection (dermis layer, superficial subcutaneous layer), application, and combined use at the hair loss position. According to different application directions, it can be referred to that the treatment is carried out once every 1 to 2 months, and 3 to 4 times for each course of treatment.

[0115] 4. Treatment evaluation criteria: 1) Average hair density; 2) Hair diameter and strength; 3) Auxiliary evaluation indicators: Epidermal thickness and hair follicle number of the scalp.

[0116] 5. Treatment conclusion:

[0117] After the treatment of patients with androgenetic alopecia (AGA) with iPSC-derived platelet-rich suspension, the hair density was significantly increased during follow-up, and the percentage increase was significantly correlated with the treatment frequency; the hair diameter increased and the results of the hair pull test decreased, proving a definite improvement in hair strength; in addition, histopathological evaluation showed that after the injection of iPSC-derived platelet-rich suspension, the epidermal thickness and the number of hair follicles in the scalp were significantly increased.

[0118] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. All the documents mentioned in the present invention are hereby incorporated by reference in their entirety in this application. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention within the spirit and principles of the present invention, and these equivalent forms of modifications also fall within the scope defined by the claims of this application.

Claims

1. A composition for beauty treatment, characterized in that, it comprises the following components: (1) Platelet concentrate prepared by differentiating pluripotent induced stem cells; (2) Additives: physiological saline of 0.9% NaCl and 5% human recombinant serum albumin; wherein, the composition can be directly injected (dermis layer, superficial subcutaneous layer), applied topically, or used in combination of injection and topical application.

2. The composition for beauty treatment according to claim 1, characterized in that, the additives further comprise 100 U / mL freeze-dried thrombin, 1% calcium gluconate and 0.05% type I collagenase.

3. A composition for beauty treatment, characterized in that, it comprises the following components: (1) Platelet concentrate prepared by differentiating pluripotent induced stem cells; (2) Additives: 10 μM ADP, 10 - 100 μg / mL fibrinogen, 2 mM CaCl 2 and 0.05 - 0.5% type I collagenase; wherein, the composition can be directly injected (into the dermis, superficial subcutaneous layer), smeared, or used in combination of injection and smearing.

4. The composition for beauty treatment according to claims 1 to 3, characterized in that, The platelet concentrate concentration is 5 to 20×10 11 / L.

5. The composition for beauty treatment according to claims 1 to 3, characterized in that, the composition is administered once every 1 to 2 months, and every 3 to 4 times is a treatment course.

6. Use of the composition according to claims 1 to 3 in the preparation of a drug for beauty treatment.

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