KR100827252B1 - Hair reproducing stimulant using autologous hair follicular cells and composition thereof - Google Patents

Abstract

A hair reproduction-stimulating composition comprising autologous hair follicular cells is provided to improve induction of hair reproduction and treatment of hair loss by using hair follicular cells which are obtained by in vitro culturing the scalp tissue through tissue culture. A hair reproduction-stimulating composition comprises autologous hair follicular cells including CD34 positive cells and CD34 negative cells which are obtained by in vitro culturing the scalp tissue in a mixing ratio of 1:1 to 1:50, preferably 1:1 to 1:25, more preferably 1:1 to 1:15 and most preferably 1:1 to 1:10, and is desirable into a form of a non-oral injection including intravenous injection, abdominal injection, intramuscular injection, subcutaneous injection and topical injection, wherein the daily dosage of the cells is 100000 to 1000000000 cells.

Description

Hair reproducing stimulant using autologous hair follicle cells and composition thereof

1 is a photograph showing a reference hair growth state of the hair growth induction score.

Figure 2 is a photograph showing the hair growth effect of each group at 14 days after the injection of hair follicle cells into each experimental group.

Figure 3 is a photograph showing the hair growth effect of each group at 21 days after the injection of hair follicle cells into each experimental group.

Figure 4 is a photograph showing the hair growth effect of each group at 28 days after the injection of hair follicle cells into each experimental group.

The present invention relates to a cell composition having hair growth inducing ability by the scalp-derived cells of positive or negative immunological properties to CD34. More specifically, the present invention relates to a hair growth-inducing cell composition, alopecia treatment agent and alopecia treatment agent containing CD34 positive cells and CD34 negative cells obtained by tissue culture of the scalp tissue in vitro at a constant mixing ratio.

Recently, as interest in beauty has increased, so has interest in the treatment of alopecia. Alopecia is a condition in which there is no hair where a normal hair should be. Hair has no important physiological function that is directly related to life, but it is very important from a cosmetic point of view as well as sun protection and hair protection. Severe hair loss can be problematic in social life and can have a serious psychological impact, which is important in terms of quality of life (Passchier J. et al ., Dermatology, 197: 217, 1998; McDonagh, AJ and Messenger , AG, Dermatol Clin., 14: 661, 1996).

Hair loss can be divided into clinically wounded alopecia and nonhairless alopecia. In the case of scarring hair loss, the hair follicles are destroyed and the hair does not come back. Hair is produced in what is called the hair follicle, and each hair follicle undergoes a phase of activity and suspension periodically (McDonagh, AJ and Messenger, AG, Dermatol Clin., 14: 661, 1996). The time intervals of the hair cycles vary depending on the body part, but in the case of the hair, the hair enters the resting period of about 3-4 months after the 26 years of growth (growth) and 2-4 weeks of degeneration. Each hair follicle will have 10 to 20 hair follicle growth cycles in its lifetime (Cotsarelis, G., et al. , Cell, 29; 61: 1329, 1990).

In normal people, the number of hairs is about 100,000, and the average length of hair growing per day is about 0.37 mm (about 1 cm a month). Normally 85-90% of the hair is in the growing phase and the number of growing hair follicles decreases with age. Therefore, 10 to 15% of hair follicles are in the degenerative or resting phase, and on average about 50 to 60 hairs fall out normally, and more than 100 hair loss days should be suspected of alopecia.

The first causes of hair loss are DHT (dihydrotestosterone), a type of male hormone, which affects hair follicles, causing protein loss and direct damage to hair follicles, leading to hair loss. It acts in conjunction with phosphorus 5α-reductase (reductase), a product of which DHT (dihydrotestosterone) affects hair follicles during DNA transcription, ie protein synthesis, hair loss due to hair follicle damage, etc. There is a theory. Second, there is genetic alopecia that is caused by the effects of male hormones by the hair loss gene, and that alopecia itself is not hereditary, and that it is inherited from ancestors, which is prone to hair loss. As the scalp develops until the age of 20 years, the skull develops before the age of 30 years, which causes thinning of the scalp, alopecia caused by skull development that causes hair loss due to blood flow disorders, and finally hair loss due to aging. Several causes have been reported (Sheen, M., Fighting Hair Loss , USA Library Publishing, Inc. p. Cm. 97-090157, 1966).

Various methods have been introduced as a treatment for hair loss, but the most commonly used surgical method is self hair transplantation (self hair transplantation) which is transplanted using its own hair. It first appeared in the 1930s and has been steadily developing since its introduction to the United States in the 1950s. In the past 15 years, various hair transplants, scalp flaps and scalp reductions in baldness have been widely practiced in North America and in Europe, particularly in France (Okuda, S., Jpn J Dermatolurol, 46: 135). , 1939).

There are only two drugs currently available: Propecia and Minoxidil, which are taken by the US Food Safety Administration. Hair loss prevention and treatment effects are reported to be effective in about 20-30% for minoxidil and 60-70% for propecia. Propecia was approved for male hair loss in the second half of 1997. Propecia contains 1 mg of a substance called finasteride, which inhibits type 2 5α-reductase (reductase). When the enzyme meets testosterone (male hormone), it turns into DHT (dihydrotestosterone), which causes hair loss. Therefore, inhibition of this enzyme can inhibit what is caused by the hair loss factor DHT hormone. In clinical photography, 66% of men who took the drug showed remarkable regrowth of hair. However, the therapeutic effect is effective at the time of administration, but when the medication is stopped, hair loss resumes as it was before taking the medicine several months later (Bouhanna, P., Dermatol Surg., 29: 1130, 2003). Thiboutot, D., Arch Dermatol., 135: 1417, 1999). Minoxidil is the first drug to be approved for the treatment of androgenetic alopecia in the second half of 1997. First of all, it was used as a treatment for hair loss in both men and women. Effective after 2 to 3 months after use, and sometimes hair falls out early, but improves over time (Bouhanna P., Dermatol Surg., 29: 1130, 2003; Messenger, AG and Rundegren J., Br J Dermatol., 150: 186, 2004). However, these medications have been reported to have many side effects such as sexual dysfunction (Messenger, AG and Rundegren J., Br J Dermatol., 150: 186, 2004).

Recently, a treatment method using genes is expected as a treatment method. In January 1998, Dr. Angela Christiano of the University of Columbia, USA, opened a new chapter in the gene therapy of hair loss by discovering and presenting genetic factors involved in systemic hair loss diseases (Ahmad W., et al. , Science 30; 279: 720, 1998). Using these gene structures, a method of delivering a desired DNA code directly to hair follicles or a treatment for blocking gene expression has been developed. However, there are so many unknowns on the efficacy, cost, stability, and future effects of such treatments, and even if the genetic factors involved in hair loss are identified, it will take a long time before the treatments using them can be realized safely. It seems.

On the other hand, a hair regeneration method has been disclosed in which the bulbs of the hair in the growing phase are extracted with the hairs attached, followed by culturing the hair follicle cells and transplanting the hair follicles into the re-extracted site, but the effect is considered to be inefficient. (US Patent No. 6,399,057).

Finally, a method for treating hair loss using stem cells is being developed. Stem cells can be obtained in four ways. The first method is to extract and grow internal cell mass in blastocyst during embryo development. The second method is to use germ cells of fetus. The third method is to make blastocyst by making embryo into ovum. As a method of obtaining, these methods are methods using an early embryo, fetus and egg. The fourth method is to obtain stem cells from an adult body. In other words, it is a method of extracting adult stem cells from some tissues of adult organs with self-renewal ability including the brain, such as extracting bone marrow cells, which is a classical method, and even cord blood can be defined as adult stem cells. In other words, adult stem cells are defined as 'cells extracted from all organs of an adult that are self-renewal, self-maintenance and multipotent that can be obtained from an adult'. To date, adult stem cells have been used in a wide range of clinical applications. For example, the identification of corneal epithelial stem cells inside the eye has led to the development of new corneal transplantation techniques, which have resulted in partial stem cell transplantation and gene therapy of autologous tissues (Cotsarelis, G. et. al. , Cell 57, 201, 1989; Tsai, RJ et al ., N. Engl. J. Med., 343, 86,2000; Bernstein, ID et al. , Blood Cells, 20:15, 1994). Other skin diseases, such as hair loss and burns, can also be expected to advance treatment in the same way as the identification of hair follicle stem cells and genetic analysis. The concept of stem cells in the epidermis has been discussed for 30 years already (Potten, CS, Cell Tiss Kinet, 7: 77, 1974). In vitro studies based on recent cell culture studies (Barrandon Y, et al ., Proc Natl Acad Sci USA, 84: 2303, 1987; Rochat, A., et al. , Cell, 76: 1063, 1994); The results of in vivo studies in the mouse show that at specific locations in the interfollicular epidermis and the upper regions of the outer root sheath of the hair follicle, or protrusions Somewhat more complex distributions and organic structures of skin stem cells have been proposed along with what are called bulb regions and similar stem cells contained in embryonic substrates of growing hair follicles (Al-Barwari, SE et al. al . , Intl J Radiat Biol , 30: 201, 1976; Morris, RJ et al . , Cancer Res 46: 3061, 1986; Cotsarelis, G. et al . , Cell , 61: 1329, 1990; Lavker, RM et al ., J Invest Derm , 101: 16, 1993; Morris, RJ, Potten CS, J Invest Derm , 112: 470, 1999; Blanpain, C. et al ., Cell , 3; 118: 635, 2004).

In terms of the interrelationship between these three segmented skin stem cells, the bulbous follicle region is thought to be the most essential part of the ultimate stem cell. In addition, hair follicle stem cells in the bulbous follicle region have been reported to express CD34 cell surface proteins, and these CD34 expressing cells have been identified as having stem cell characteristics (Trempus, CS et al. , J) . Invest Dermatol, 120: 501, 2003).

However, no clear culture of hair follicle stem cells has been established, and markers of hair follicle stem cells are unclear. In addition, although it is known that hair follicle stem cells exist in some hair follicles, a large amount of stem cells are required for the actual treatment of human baldness, but the stem cells are still proliferated to the extent that can be applied clinically. The technique to make was insignificant. Stem cell marker proteins have not yet been accurately established, and hair loss treatment using stem cells is still insufficient.

Therefore, the present inventors have made efforts to develop new hair growth induction and hair loss treatment method using hair follicle cells. As a result, the scalp tissues were cultured in vitro, and the CD34 positive cells and CD34 negative cells derived from the tissues were separated. The present invention was completed by mixing the cells in a certain ratio and confirming the hair growth efficacy in animal experiments.

It is an object of the present invention to provide a cell composition for inducing hair growth comprising a predetermined mixing ratio of CD34 positive cells and CD34 negative cells obtained by tissue culture of the scalp tissue in vitro.

Another object of the present invention to provide a hair loss treatment agent and alopecia treatment agent containing the CD34 positive cells and CD34 negative cells as an active ingredient.

In order to achieve the above object, the present invention provides a cell composition for inducing hair growth comprising a constant mixing ratio of CD34 positive cells and CD34 negative cells obtained by tissue culture of the scalp tissue in vitro.

The present invention provides a hair loss treatment agent and alopecia treatment agent containing the CD34 positive cells and CD34 negative cells as an active ingredient.

Hereinafter, the present invention will be described in detail.

In one aspect, the present invention relates to a cell composition for inducing hair growth comprising CD34 positive cells and CD34 negative cells obtained by culturing the scalp tissue in vitro as an active ingredient.

'CD34 positive cells' and 'CD34 negative cells' of the present invention include stem cells having the characteristics of stem cells, that is, undifferentiated, infinite proliferation and differentiation into specific cells, and can be applied to the present invention regardless of origin have. Stem cells can be extracted from the internal cell mass of the blastocyst during embryonic development, embryonic stem cells that are capable of differentiating into a variety of cells, and can be extracted from specific tissues of the adult and differentiated into specific derived tissues. Adult stem cells with their limited capacity to be divided into two, which can be obtained from all tissues, especially from bone marrow, blood, liver, skin, intestine, pancreas, brain, skeletal muscle and pulp. can do. At this time, in order to apply to the hair growth induction in the present invention, it is preferable that the adult stem cells are separated from the scalp including the hair follicles, more preferably autologous scalp derived adult stem cells. However, the present invention is to establish a hair growth induction composition by analyzing the hair growth ability of the CD34 positive and CD34 negative cells and cells mixed with them at a certain ratio.

In another aspect, the present invention relates to a hair loss treatment agent and a hair loss treatment agent containing CD34 positive cells and CD34 negative cells as an active ingredient.

In the present invention, 'hair growth induction' refers to the ability to induce hair follicles to form hair follicles in the hair loss site or hair loss site.

The hair composition-inducing cell composition, the hair loss treatment agent and the alopecia treatment agent of the present invention is preferably prepared in a state in which CD34 positive cells and negative cells are mixed.

CD34 positive cells and CD34 negative cells contained in the hair composition-inducing cell composition, the hair loss treatment agent and the alopecia treatment agent of the present invention may exhibit the maximum hair growth induction effect only when mixed with a certain ratio. Therefore, the mixing ratio of the two cells may be 1: 1 to 1:50, preferably 1: 1 to 1:25, more preferably 1: 1 to 1:15, and most preferably 1: 1 to 1:10. A single dose may contain from 10 ⁴ to 10 ⁸ cells per day, but the amount may be determined by a person skilled in the art and may be determined by factors such as the hair loss and response sensitivity of the recipient. It can be prescribed in various ways. In addition, the number of administrations may be administered once or several times depending on the condition of the subject.

The parenteral administration including the intravenous administration, the intraperitoneal administration, the intramuscular administration, the subcutaneous administration, or the topical administration is preferred, and more preferably subcutaneous administration. Alternatively, it may be administered by topical administration, and may be administered by injecting directly into a site requiring hair loss or hair growth.

Hair composition-inducing cell composition, hair loss treatment agent and alopecia treatment agent of the present invention can be applied to the scalp for the treatment of typical male alopecia and so-called alopecia that can occur after menopause or ovarian removal surgery, which is recognized as 'baldness'. It can be applied to any part of the body that needs hair growth. For example, it can be used to improve the condition of hair damaged by trauma scar, wide forehead or M forehead, lashes or eyebrows and alopecia for the purpose of simple cosmetic effect.

Pharmaceutically acceptable carriers included in the cell composition for inducing hair growth of the present invention, the alopecia treatment agent and the alopecia treatment agent are commonly used in the preparation, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, Calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and minerals Oils and the like, but are not limited thereto.

The cell composition for inducing hair growth, the hair loss treatment agent and the alopecia treatment agent of the present invention may further include a lubricant, a humectant, an emulsifier, a suspension agent, a preservative, and the like, in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

The cell composition for inducing hair growth, the alopecia treatment agent and the alopecia treatment agent of the present invention may be prepared using a pharmaceutically acceptable carrier and / or excipient according to a method which can be easily carried out by one of ordinary skill in the art. It can be prepared in unit dosage form or by incorporation into a multi-dose container, and may further include a dispersing agent or stabilizer.

The hair composition-inducing cell composition, the hair loss treatment agent and the alopecia treatment agent of the present invention may be used as a single therapy, but may also be used in combination with other conventional hair growth-inducing drug therapy or surgical therapy, and maximized when the combination therapy is performed. Efficacy may be exhibited.

Drug formulations that can be used with the hair composition-inducing cell composition, hair loss treatment and alopecia treatment of the present invention are male hormones or DHT-inducing inhibitors including finasteride, dutasteride, thioctol and RU58841, parathyroid Hormone (PTH) inhibitors, minoxidil, Busserelin (Burserelin), blood circulation improvers and compositions applied to mesotherapy, and the like, and surgery includes hair transplantation, scalp flap surgery, scalp reduction and the like.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention. .

Example 1 Culture of Scalp-derived Hair Follicle Cells

After scalp-derived tissue was cut, medium containing collagenase type IA (10% fetal bovine serum (Gibco, USA), 100 units of penicillin, 0.1 mg / ml streptomycin, 0.25 μg / ml neomycin ( Gibco, USA), DMEM containing 100 μg / ml normosin (Invivogen, USA) and medium containing 2 mg / ml collagenase type IA (Sigma, USA)) Chemical degradation was allowed to occur in a gravity convection incubator for 30 minutes at 37 ° C., m / min. The chemically digested tissue was collected by centrifugation and washed three times with PBS, followed by M199 / F12 serum medium (M199 and F12 at 1: 1, 0.62 μg / ml insulin (Sigma, USA), 0.62 μg / ml transferrin). (Gibco, USA), 100 units of penicillin, 0.1 mg / ml streptomycin, 0.25 μg / ml neomycin (Gibco, USA), 10 ng / ml rEGF (Sigma, USA), 10 ng / ml bFGF (Sigma, USA), 100 μg / ml normosin (Invivogen, USA), 10% fetal bovine serum and 1 mM N-acetyl-L-cysteine (Sigma, USA)) . After about three days, when tissues began to adhere to the bottom of the culture vessel, the cells were replaced with M199 / F12 serum-free medium and cultured for one week, and keratinocyte-free medium without normosin (0.2 mM ascorbic acid (Sigma, USA)). Cells 1 (groups of cells attached to the bottom of the cell culture vessel from the beginning of the culture) and cells 2 depending on the nature of the culture medium by replacing them with defined serum keratinocytes serum-free medium (Gibco cat # 10785-012, USA). (Cell group attached after passage 1) and cell 3 (floating cell group), respectively.

Example 2 Classification of Scalp-derived Hair Follicle Cells by MACS

CD34 positive cells were isolated using a commercially available MACS (Magnetic Cell Sorting, Miltenyi Biotec Inc., Germany). The cells obtained in Example 1 were harvested, washed, and then suspended in single cells by pipetting by adding medium, and 150-300 μl MACS buffer, 50-100 μl blocking regent, and 50-100 μl microbeads were added sequentially. . The process of preparing the mixed solution was carried out in the dark state, the tube containing the mixed solution was wrapped in aluminum foil, and reacted at 4 ℃ for 30 minutes to 4 hours. Thereafter, the aluminum foil was peeled off, and about 10 times the volume of the MACS buffer of the mixed solution was added to the tube, mixed well by a pipette, centrifuged at 1200 rpm for 5 to 6 minutes, and the supernatant was removed. 500 μL MACS buffer was added according to the number of cells, and then CD34 positive cells and CD34 negative cells were separated using MACS according to the manufacturer's recommended method. At this time, the number of collected CD34 positive cells was about 1 to 2 x 10 ^7, and the cells were resuspended in 100 μl of sterile physiological saline to be used for further animal experiments.

Example 3 Hair Growth Treatment Effect of Scalp-derived Hair Follicle Cells

Six-week-old male BALB / cAnNCrj Bgi-nu nude mice (Central experimental animals) were injected subcutaneously with 1 × 10 ⁵ CD34 positive cells. The control group was photographed before administration and compared with the change in the state of the mice after time. The 67 nude mice used in this experiment were bred in the SPF area of the Seoul National University Veterinary Medicine Laboratory. The experiment was performed by separating the groups so that each of them was randomly included. The composition of the test group is shown in the table below (Table 1).

group Cell count Marisu Control No Action (CS) NA 5 Saline administration (CN) NA 5 CD34 positive group Low capacity (NL) 1X 10 ⁴ 5 Medium capacity (NM) 1X 10 ⁵ 5 CD34 single voice group Low capacity (PL) 1X 10 ⁴ 5 Medium capacity (PM) 1X 10 ⁵ 5 CD34 positive / CD34 negative mixed group 2: 1 (T1) 6.6X10 ⁴ : 3.3X10 ⁴ 5 1: 1 (T2) 5X10 ⁴ : 5X10 ⁴ 5 1: 2 (T3) 3.3X10 ⁴ : 6.6X10 ⁴ 5 1: 4 (T4) 2X10 ⁴ : 8X10 ⁴ 5 1: 8 (T5) 1.1X10 ⁴ : 8.8X10 ⁴ 5

In addition, by using a commercial hair loss treatment agent minoxidil (Minoxidil, Pfizer, USA) in combination with the cultured cells to include three mice in each group to evaluate the hair growth effect was prepared in the experimental group compared to Table 1 (Table 2 ).

group Cell count Marisu Minoxidil Control (MC) NA 3 Minoxidil kill group CD34 positive single group (MP) 1X 10 ⁵ 3 CD34 negative single group (MN) 1X 10 ⁵ 3 CD34 positive / CD34 negative mixed group (MT) 5X10 ⁴ : 5X10 ⁴ 3

One day before the hair follicle cells were administered to the experimental group, the hairs of the experimental animals were removed with calcium thioglycorate (Sigma, USA), and the scalp of the experimental animals was observed for 30 days. The hair growth induction scores were divided and recorded daily, and the body weight of the animals was measured before and at the beginning of the experiment, and an autopsy was performed at 31 days after the experiment.

Hair induction score was evaluated from 0 to 3 points according to the state of hair growth by visual criteria (Fig. 1). In addition, one-way ANOVA was performed for statistical analysis of the data on the condition of each experimental animal, and if the significance was recognized, Dunnett's t-test was performed for statistical significance between the control group and the experimental group. (p <0.05).

According to the hair growth induction score recorded after the administration to the experimental animals in each group as shown in Table 1, the hair growth efficacy was most maximal after the 24th day of the experiment, the hair growth initiation effect was the fastest in the T3 group, the hair follicle cell mixing group. In addition, the minoxidil and the mixed administration (MT) showed a similar effect, but the efficacy after 24 days showed that T3 is superior, and the hair growth effect on the 14th, 21st and 28th day after hair follicle cell administration (Figures 2, 3 and 4). In addition, the chart of calculating the average value of each group by describing the hair growth induction score for each individual up to 30 days when the day of cell administration was 0 days is as follows (Table 3).

DAY 00 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 CN 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0.2 0.4 0.6 0.6 0.4 0 CS 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0.2 0.2 0.6 0.6 0.2 0.2 NL 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0.2 0.4 0.8 0.8 0.4 0.4 NM 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0.4 0.8 1.2 1.2 0.8 0.8 PL 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0.4 0.6 0.6 0.8 0.6 PM 0 0 0.2 0.4 0.6 One 0.8 0.8 0.8 0.8 0.8 1.2 1.2 1.2 1.2 0.8 1.6 2 1.6 0.8 0 T1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0.2 0.4 0.4 0.6 0.4 0.4 T2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0.6 One 1.4 One One One T3 0 0.6 0.8 One One One 0.8 0.8 0.6 0.6 0.4 0.6 0.6 0.6 0.6 2.2 2.8 2.4 2.2 One 0.2 T4 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0.4 1.4 2 1.2 0 0 T5 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 One 1.4 1.8 0.6 0.2 0.2 MC 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0.2 0.2 0 0 MN 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0.2 0.6 0.6 0.6 0.2 0.2 MP 0 0 0 0 0 0 0 0 0 0 0.2 0.4 0.4 0.4 0.4 0.2 0.4 0.4 0.4 0.6 0.4 MT 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0.2 0.6 0.8 0.2 0.2 0

As described in detail above, the present invention relates to a cell composition for inducing hair growth, alopecia treatment agent and alopecia treatment agent containing CD34 positive cells and CD34 negative cells obtained by culturing the scalp tissue in vitro in a constant mixing ratio. The cell composition for inducing hair growth thus obtained is very useful as a novel cell therapy for promoting hair growth.

delete

Having described the specific part of the present invention in detail, it is apparent to those skilled in the art that the specific technology is merely a preferred embodiment, and the scope of the present invention is not limited thereto. Therefore, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

Claims (9)

delete delete delete A hair growth cell composition containing CD34 positive cells and CD34 negative cells in a ratio of 1: 2 obtained by tissue culture of the scalp tissue in vitro. delete delete A hair loss treatment agent comprising the cell composition for hair growth of claim 4 as an active ingredient. delete A hair loss treatment agent containing the hair growth cell composition of claim 4 as an active ingredient.

Images