CN101251528A - Method for evaluating or screening hair growth-regulating agent - Google Patents
Method for evaluating or screening hair growth-regulating agent Download PDFInfo
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Abstract
The present invention provides a method for evaluating or screening a hair growth-regulating agent which utilizes a readily available animal. The present invention is directed to a method for evaluating or screening a hair growth-regulating agent which comprises subjecting hair follicles of a pig to an organ culture in the presence of a test substance, and evaluating or selecting a substance which promotes or suppresses the growth of the hair follicles.
Description
Technical field
The present invention relates to utilize the hair follicle evaluation of pig or the method for screening hair growth-regulating agent.
Background technology
Usually, about hair, people often wish to promote the hair growth or suppress the hair growth according to the position.For example, many hypotrichosis and trichomadesiss that artificially cause because of stress and heredity etc. feel worried.In addition, the somebody handles the useless hair of what is called and feels trouble.
About hair, known existence depends on the hair of male sex hormone and does not rely on the hair of male sex hormone.For example, think: the hair of the male pattern alopecia disease that the alopecia from frons to head or this part are drawn back and beard and shank-feathering, armpit hair etc. is closely related with male sex hormone, and temple portion hair, back of head hair, eyebrow, eyelashes then are subjected to male sex hormone to influence little (non-patent literature 1, non-patent literature 2).
Therefore, the hair growth promoter and the hair growth inhibitor (patent documentation 1 and 2) of the elongation that is used to regulate hair have been developed in all its bearings.
Based on the viewpoint of can be rapidly and estimating easily or screening, in recent years, in the exploration of hair growth promoter and hair growth inhibitor, the cellular incubation of the hair follicle of employing people and mouse rat and organ culture method's (non-patent literature 3～5, patent documentation 3 and 4) of hair follicle.The screening of adopting the organ culture of people's hair follicle to carry out, though the reliability height of data, but people's hair follicle itself be difficult to obtain, so be unsuitable for carrying out a large amount of screenings.In addition, the screening that the organ culture of the hair follicle of employing mouse rat is carried out though can often obtain, owing to use animal used as test, based on zoophilous viewpoint, also is unsuitable for screening in a large number.
[patent documentation 1] TOHKEMY 2005-206536 communique
[patent documentation 2] TOHKEMY 2006-8657 communique
[patent documentation 3] Japanese kokai publication hei 11-49647 communique
[patent documentation 4] TOHKEMY 2002-62289 communique
People such as [non-patent literature 1] Ebling FJ, clinical endocrine metabolism (Clin EndocrinolMetab.), 15 volumes, 319-339 (1986)
People such as [non-patent literature 2] Stenn KS, physiology summary (Physiol Rev.), 81 volumes, 449-494 (1993)
People such as [non-patent literature 3] T.Jindo., dermatology magazine (The Journal ofDermatology), Vol.20,756-762,1993.
[non-patent literature 4] Yu mound really and ▲ play pool thousand to add day skin will, 104 (8), 979-987,1994.
People such as [non-patent literature 5] M.P.Philpott, cell science magazine (Journal of CellScience), 97,463-471,1990.
Summary of the invention
The present invention relates to provide the hair follicle evaluation of the animal that utilization obtains easily or the method for screening hair growth-regulating agent.
The present inventor finds after having studied the system of new evaluation or screening hair growth regulatory substance, when the hair follicle with each position of pig carries out organ culture, exist its growth be subjected to 5 α-dihydrotestosterone (below, be abbreviated as DHT) influence hair follicle and the hair follicle that not influenced by DHT, by using these hair follicles, can estimate or screen the growth regulator of male sex hormone dependence or dependent/non-dependent hair.
That is, the invention provides a kind of the evaluation or the method for screening hair growth-regulating agent, it is characterized in that: in the presence of analyte, the hair follicle of pig is carried out organ culture, the material that promotes or suppress the growth of this hair follicle is estimated or selected.
The present invention also provides a kind of method of estimating or screening male sex hormone dependence hair growth-regulating agent, it is characterized in that: in the presence of analyte, the belly of pig or the hair follicle at back are carried out organ culture, the material that promotes or suppress the growth of this hair follicle is estimated or selected.
The present invention also provides a kind of method of android type alopecia with the hair growth promoter of estimating or screen, it is characterized in that: in the presence of analyte, the hair follicle of the belly of pig is carried out organ culture, the material of the growth that promotes this hair follicle is estimated or selected.
The present invention also provides a kind of method of estimating or screening male sex hormone dependence hair growth inhibitor, it is characterized in that: in the presence of analyte, the hair follicle at the back of pig is carried out organ culture, the material of the growth that suppresses this hair follicle is estimated or selected.
The present invention also provides a kind of method of estimating or screening male sex hormone dependent/non-dependent hair growth-regulating agent, it is characterized in that: in the presence of analyte, the hair follicle of flank, pars inguinalis, shoulder or the buttocks of pig is carried out organ culture, the material that promotes or suppress the growth of this hair follicle is estimated or selected.
According to the present invention, because can use the hair follicle of the porker that obtains in a large number easily, so can estimate or select to have the material of mao growth-promoting effect or hair growth inhibitory effect easy and effectively.
Embodiment
The method of evaluation of the present invention or screening hair growth-regulating agent is that the hair follicle with the privileged site of pig carries out organ culture and to promoting or suppressing the method that the material of its growth is estimated or selected.
Shown in following embodiment, in the presence of as the DHT of one of male sex hormone, the hair follicle at each position of pig is carried out organ culture, when with the elongation of the hair of the same area of not adding DHT relatively during the growth of this hair follicle, find: the elongation of the hair of belly reduces significantly, the elongation of the hair at back increases significantly, and the elongation of each mao of flank, pars inguinalis, shoulder or buttocks is (embodiment 1) about equally.That is, in the growth of the hair follicle of pig, find:, exist its growth to be subjected to hair follicle that DHT influences and the hair follicle that not influenced by DHT according to the position, source of the hair follicle of pig.
On the other hand,, just know a long time ago about people's hair: the android type alopecia that the alopecia from frons to head or this part are drawn back, its symptom develops with male sex hormone, and known antiandrogen agent is effective to this.In addition, it was reported, the growth of beard and shank-feathering is subjected to male sex hormone to promote (people such as Ebling FJ, Clin Endocrinol Metab., 15 volumes, 319-339 (1986)), and think that temple portion hair, back of head hair, eyebrow, eyelashes are subjected to male sex hormone to influence little (people such as Stenn KS, Physiol Rev., 81 volumes, 449-494 (2001)).
Therefore think that the growth of the hair follicle of the belly of pig is that male sex hormone is dependent and be similar to the growth of the hair that is suppressed by male sex hormone, for example the growth of people's hair etc. from frons to head; The growth of the hair follicle at the back of pig, be male sex hormone dependent and be similar to by male sex hormone promote the hair growth, for example growth of people's beard, shank-feathering, chesk hair, pubes etc.; The growth of the hair follicle of the hair follicle of the flank of pig, the hair follicle of pars inguinalis, shoulder and the hair follicle of buttocks is similar to the hair that not influenced by male sex hormone, for example the growth of the hair of the hair of people's temple portion, back of head, eyebrow, eyelashes etc.
Thereby, think and can estimate or screen male sex hormone dependence hair growth-regulating agent, android type alopecia hair growth-regulating agent with the index that is grown to of the hair follicle of the privileged site of pig with a hair growth promoter, male sex hormone dependence hair growth inhibitor and male sex hormone dependent/non-dependent hair growth-regulating agent etc.
Particularly, can carry out evaluation of the present invention or screening by for example following operation (1)～(3).
(1) makes the operation of hair follicle contact analyte of the privileged site of pig;
(2) operation of this hair follicle being carried out organ culture and measuring its growth and will grow and compare with the growth of the hair follicle that does not contact analyte;
(3) will promote or suppress the operation that the analyte of the growth of hair follicle is estimated or selected as hair growth-regulating agent based on the result of (2).
The hair follicle of so-called pig, be meant the skin of taking from pig around all tissues of hair root, refer to comprise all tissues of relevant organ such as the elongation with hair of hair shaft, papilla, hair matricyte, hair root etc. particularly.
Among the present invention, in this hair follicle, use the hair follicle (hair follicle in growth period) that is in growth period.
Here, take the pig of object, so long as the pig of hairiness can be used any kind for becoming hair follicle.Yet, usually, can use to reaching meat purpose and hybridize the kind of the pig that obtains.
As meat and modified pig, for example, can enumerate the Yorkshire kind (having large, medium and small 3 types) of area, the state, Yorkshire of Britain being used the adularescent hair that land race and Chinese variety, Naples kind (Neapolitan breed) and the mating such as (Lester breed) of Leicester's kind obtain; Berkshire (Berkshire) and the land race in area, Weir summer prefecture (Wilshire) and the Berkshire kind that the black hair is arranged that hybridization such as Siam's kind (Siamese breed), Chinese variety and Naples kind obtain with Britain; Lan Deruisi (land race) kind of the adularescent hair that the land race of Denmark and big Yorkshire kind mating are obtained; Originate in the Massachusetts of the U.S. and the hampshire kind that the black hair is arranged of the Kentucky State; Du Luoke (Duroc) kind that obtains by the pig of the red colour system that originates in Europe and pig mating in the U.S. etc. at the red colour system of New Jersey after improvement.
As in Japan by the registration kind that the blood lineage keeps, middle Yorkshire kind (Y), Berkshire kind (B), Lan Deruisi kind (L), big Yorkshire kind (W), hampshire kind (H), these 6 kinds of Du Luoke kind (D) are arranged.As general porker, except the black pig of purebred Berkshire kind, also have a lot of hybrids for these kind combinations with one another are obtained.For example, the pig " Tochigi wood L " that can enumerate the blood lineage of Lan Deruisi kind (L) gives birth to " Tochigi wood LaLa pig " again with Du Luoke kind (D) mating with " the LW sow " that big Yorkshire kind (W) mating obtains; " Tokyo X " that Pig Beijing Black kind and Berkshire kind and Du Luoke mixing breed are obtained etc.
Wherein, the kind and the hybrid thereof of the adularescent hair of the Yorkshire kind of the easy judgement of preferred use hair in growth period, Lan Deruisi kind etc.When using porker, can obtain hair follicle from the skin of having obtained the pig after the meat, can only not injure animal for obtaining hair follicle, be preferred therefore.
As using the position, can enumerate belly, back, flank, pars inguinalis, shoulder or buttocks etc., can select according to estimating purpose.That is: for of evaluation or the screening of android type alopecia, preferably use the hair follicle of belly with the hair growth promoter; For the evaluation or the screening of growth regulating (the hair growth promotes or the hair growth inhibited) agent of the male sex hormone dependence hair of beard and arm hair shank-feathering etc., preferably use the hair follicle at back; For the evaluation or the screening of the growth regulating of male sex hormone dependent/non-dependent hair (the hair growth promotes or the hair growth inhibited) agent, preferably use the hair follicle of flank, the hair follicle of pars inguinalis, the hair follicle of shoulder or the hair follicle of buttocks.
Can adopt following method to carry out taking of hair follicle, promptly cut pigskin, under aseptic condition, behind the excision adipose tissue, separate hair follicle corresponding to the position of estimating purpose.
Can adopt and for example add analyte in advance and in nutrient solution, make after its concentration that reaches regulation the method for mounting hair follicle in nutrient solution, perhaps, add analyte and in mounting has the nutrient solution of hair follicle, make its method that reaches the concentration of regulation, carry out contacting of hair follicle and analyte.
Can adopt the organ culture method of known hair follicle of organ culture (non-patent literature 4) etc. of hair follicle of the beard of mouse, carry out the organ culture of hair follicle.For example: on the dish of sterilization net that can be by nutrient solution is arranged in infiltration etc., the hair follicle that mounting is taked under 37 ℃ temperature, is containing CO
2The air gas phase under, be generally 2～56, be preferably 4～21 days cultivation.
As nutrient solution, for example, can enumerate RPMI1640 nutrient culture media, William ' s MediumE nutrient culture media, DMEM/HamF12 (1: 1) nutrient culture media etc.Can suitably add agar and gelatin.In addition, as required, also can add microbiotic, amino acid, serum, growth factor, live body extract etc.
The mensuration of the growth of hair follicle, for example, can enumerate the method for utilizing image analysis or measuring the increase and decrease of the elongation of hair and thickness at microscopically with micrometer, the cell-proliferation activity in the hair follicle after the mensuration organ culture or the method for proliferative cell quantity, the intensity of the growth of mensuration and hair follicle is more common in the method for the apoptotic cell in the hair follicle etc. inversely.
In the mensuration of the growth of hair follicle, adopt image analysis or when microscopically is measured the method for increase and decrease of the elongation of hair and thickness with micrometer, for example, can enumerate mensuration, the elongation of the hair when measuring then from the cultivation beginning by the length of ball top portion basal part to the hair shaft top; Perhaps the thickness of the hair shaft by near the elongation relatively high-quality green tea and the hair root is measured the method etc. of the increase and decrease of thickness.
Under the above-mentioned situation, the evaluation of the material of the growth of promotion or inhibition hair follicle can be as described below, even hair follicle contact analyte, carry out organ culture, with mensuration obtain the hair elongation or thickness with do not contact analyte mao elongation or thickness (reference standard) compare, try to achieve the length growth rate (%) when the regulation reference standard is 100 or the rate of change (%) of thickness, estimate based on this result.And, be worth highly more, estimate the growth that analyte promotes hair more; Perhaps value is low more, estimates the growth that analyte suppresses hair more.
In addition, when measuring the proliferation activity, respiratory activity of the cell in the hair follicle or proliferative cell quantity, for example, thus can be set forth in and add bromodeoxyribouridine in the nutrient culture media of organ culture and according to the synthetic method that can measure cell-proliferation activity of the quantitative determination DNA that imports the bromodeoxyribouridine among the DNA; Perhaps, use Alamar Blue to detect and MTT detect (with mitochondrial NADH reduction as the tetrazolium salts of matrix interpolation and measure the absorbance of its product and the method for trend) etc. the method for respiratory activity of mensuration cell; Perhaps, method of proliferative cell quantity etc. is measured in the dyeing of being undertaken by the antibody as the Ki67 of proliferative cell mark and PCNA (proliferating cell nuclear antigen, Proliferation Cell Nuclear Antigen).
In the above-mentioned situation, the evaluation of the material of the growth of promotion or inhibition hair follicle can be, make hair follicle contact analyte, carry out organ culture, proliferation activity or the proliferative cell quantity measured are as mentioned above compared with the proliferation activity or the proliferative cell quantity (reference standard) of the hair follicle that does not contact analyte, try to achieve proliferation activity ratio (%) or proliferative cell quantity ratio (%) when the regulation reference standard is 100, estimate based on this result.And the value of calculating gained is high more, estimates the growth that analyte promotes hair more; Perhaps, the value of calculating gained is low more, estimates the growth that analyte suppresses hair more.
In addition, when measuring the apoptotic cell in the hair follicle, for example, can enumerate the method for the sheet segment DNA amount when measuring Apoptosis and carrying out; Perhaps, measure the method etc. of apoptotic cell quantity according to TUNEL method etc.
In the above-mentioned situation, the evaluation of the material of the growth of promotion or inhibition hair follicle can be: make hair follicle contact analyte, carry out organ culture, sheet segment DNA amount or the apoptotic cell quantity measured are as mentioned above compared with the sheet segment DNA amount or the apoptotic cell quantity (reference standard) of the hair follicle that does not contact analyte, try to achieve sheet segment DNA amount ratio (%) or apoptotic cell quantity ratio (%) when the regulation reference standard is 100, estimate based on this result.And the value of gained is low more, estimates the growth that analyte promotes hair more; Perhaps, the value of gained is high more, estimates the growth that analyte suppresses hair more.
Embodiment
Embodiment 1
(1) separation of hair follicle
As pigskin, the skin that use obtains from the pig of the hybrid of the adularescent hair of raising as porker in Japan, cut by appropriate size, excise unnecessary fat, under aseptic environments, in two chlorobenzene biguanides ethane liquid (5% pair of chlorobenzene biguanides ethane liquid (manufacturing of Sumitomo drugmaker) is diluted with water to 5～20 times), soaked 5 minutes～10 minutes, carry out disinfection, then, carry out several with D-PBS and clean.
Then, under stereomicroscope, with pincet and scalpel (FEATHER No 10), separate hair follicle, it is concentrated in the nutrient culture media from the pigskin of having handled.As nutrient culture media, use RPMI1640 nutrient culture media (Gibco cat.No.11835-030), William ' s Medium E nutrient culture media (Gibco cat.No.1255 1-032) and DMEM/HamF12 (1: 1) nutrient culture media (Gibco cat.No.11039-021).
(2) DHT is to the influence of the hair follicle organ culture of pig
According to the method for (1), obtain the hair follicle of pig from back, flank, belly, pars inguinalis, buttocks, the shoulder of pig.The hair follicle that separation obtains is put into 24 well culture plates, and every hole is put into 1.In every hole, add nutrient culture media William ' s Medium E nutrient culture media (Gibco cat.No.12551-032) 400 μ L.Use is added with the nutrient culture media of penicillin/streptomycin (Gibco cat.No.15140-122) solution in 1% ratio.
Use above-mentioned William ' s Medium E nutrient culture media, being modulated into and making the ultimate density of DHT (Sigma cat No.A-8380) in nutrient culture media is 10ng/mL.At 37 ℃, 5%CO
2Environment under, in constant temperature oven, carry out cultivating in 6th.
Every 1 day or displacement 1 subculture on the 2nd.The elongation and the reference standard of the hair of cultivation after 6 days are compared.
Standard is used William ' the s Medium E nutrient culture media that does not add DHT in contrast, and the elongation of the hair after the regulation organ culture is 100%.
Moreover being determined as of elongation of hair: it day was the 0th that regulation is cultivated beginning, on the 0th and the 6th, utilize ccd video camera (pixera model.No.PVC 100C), take the MIcrosope image of virtual condition,, measure by the length variations of ball top portion basal part to the hair shaft top according to this image.
[table 1]
| Hair follicle is taked the position | DHT(10ng/mL) * |
| The back | 179％ |
| Flank portion | 108％ |
| Belly | 75％ |
| Pars inguinalis | 102％ |
| Shoulder | 99％ |
| Buttocks | 102％ |
*Organ culture the 6th day, the regulation reference standard is 100%.
Show that by table 1 even the hair follicle of pig, its growth also is subjected to the influence of DHT; Also show according to the position influence degree difference of DHT.
The hair follicle of belly, its growth is suppressed by DHT.Therefore, the elongation promoter of this hair follicle can become the effective hair of android type alopecia growth promoter.
In addition, the hair follicle at back, its growth is promoted by DHT.Therefore, the elongation inhibitor of this hair follicle can become the male sex hormone dependence and be promoted the hair growth inhibitor of the hair of growth by male sex hormone.
The hair follicle at other position is subjected to the influence of DHT little.Therefore, the elongation promoter of this hair follicle or the elongation inhibitor, can become do not work via the male sex hormone effect the hair the hair growth promoter or the hair growth inhibitor.
The evaluation of embodiment 2 hair growth-regulating agents
(1) t-flavanones and TGF-β 2
According to operation similarly to Example 1, obtain the hair follicle of the buttocks of pig.As nutrient culture media, use RPMI1640 (Gibco cat.No.11835-030) nutrient culture media.Use is added with the nutrient culture media of penicillin/streptomycin (Gibco cat.No.15140-122) solution in 1% ratio in the RPMI1640 nutrient culture media.
Use above-mentioned RPMI1640 nutrient culture media, modulation promptly is identified the t-flavanones (trans-3 with mao growth-promoting effect as the effective constituent of the hair growth promoter that does not belong to pharmaceuticals in the organ culture of the hair follicle of the beard of rat, 4 '-dimethyl-3-hydroxyl flavanones (people such as hole field, fragrance magazine (Fragrance Journal), the 31 the 2nd phase of volume, 33-40 (2003))), making the ultimate density in the nutrient culture media is 1 μ M (about synthetic method, with reference to TOHKEMY 2000-198779).And, have mao Growth Inhibition TGF-β 2 (R﹠amp about in the organ culture of people's hair follicle, being identified; D System, Inc., model: 102-B2-001/CF) (people such as Philpott, JCell Sci, 97 volumes, 463-47 (1990); People such as Soma, J Invest Dermatol, 118 volumes, 993-997 (2002)), also be modulated into 10ng/mL.
At 37 ℃, 5%CO
2Constant temperature oven in, carry out cultivating in 8th.Every 1 day or displacement 1 subculture on the 2nd, measure the elongation and the reference standard of cultivating the hair after 8 days and compare.
Adopt the increment of method evaluation hair follicle similarly to Example 1.
Standard is used the RPMI1640 nutrient culture media that does not add analyte in contrast, and the elongation of the hair after the regulation organ culture is 100%.
[table 2]
| Analyte | The regulation reference standard is 100 o'clock a hair length growth rate (%) |
| T-flavanones (1 μ M) | 145 |
| TGF-β2(10ng/mL) | 59 |
Organ culture the 8th day, the RPMI1640 nutrient culture media
Compare with reference standard, seen that the t-flavanones has mao length growth rate and reaches 145% hair elongation facilitation effect, and seeing aspect the TGF-β 2 that having mao length growth rate is that 59% hair elongation suppresses effect.
(2) ring sporidesmin A
According to operation similarly to Example 1, obtain the hair follicle of the buttocks of pig.As nutrient culture media, use William ' s E nutrient culture media (Sigma cat.No.W1878) nutrient culture media.Use is added with the nutrient culture media of penicillin/streptomycin (Gibco cat.No.15140-122) solution in 1% ratio in William ' sE nutrient culture media.
Use above-mentioned William ' sE nutrient culture media, be modulated in the organ culture of people's hair follicle and be identified ring sporidesmin A (CsA) (Sigma cat.No.C-1832) (people such as Taylor with mao growth-promoting effect, J Invest Dermatol, 100 volumes, 237-239 (1993)), making the ultimate density in the nutrient culture media is 100ng/mL, 1000ng/mL.
At 37 ℃, 5%CO
2Constant temperature oven in, carry out cultivating in 7th.Every 1 day or displacement 1 subculture on the 2nd, measure the elongation of cultivating the hair after 7 days, and compare with reference standard.
Adopt the increment of method evaluation hair follicle similarly to Example 1.
Standard is used the William ' s E nutrient culture media that does not add analyte in contrast, and the elongation of the hair after the regulation organ culture is 100%.
[table 3]
| Analyte (ng/mL) | The regulation reference standard is 100 o'clock a hair length growth rate (%) |
| CsA(100ng/mL) | 116 |
| CsA(1000ng/mL) | 120 |
| Organ culture the 7th day, William ' s E nutrient culture media | |
Compare with reference standard, seen that CsA (100ng/mL) has mao length growth rate and reaches 116% hair elongation facilitation effect, and seen aspect the CsA (1000ng/mL) that having mao length growth rate reaches 120% hair elongation facilitation effect.
(3) sum up
Therefore, show: can be that index is estimated hair growth-regulating agent with the elongation of the hair of pig buttocks.
Claims (5)
1. estimate or the method for screening hair growth-regulating agent for one kind, it is characterized in that:
In the presence of analyte, the hair follicle of pig is carried out organ culture, the material that promotes or suppress the growth of this hair follicle is estimated or selected.
2. estimate or the method for screening male sex hormone dependence hair growth-regulating agent for one kind, it is characterized in that:
In the presence of analyte, the belly of pig or the hair follicle at back are carried out organ culture, the material that promotes or suppress the growth of this hair follicle is estimated or selected.
3. estimate or screening is used for the method for the hair growth promoter of android type alopecia for one kind, it is characterized in that:
In the presence of analyte, the hair follicle of the belly of pig is carried out organ culture, the material of the growth that promotes this hair follicle is estimated or selected.
4. estimate or screening is used for the method for the hair growth inhibitor of male sex hormone dependence hair for one kind, it is characterized in that:
In the presence of analyte, the hair follicle at the back of pig is carried out organ culture, the material of the growth that suppresses this hair follicle is estimated or selected.
5. estimate or the method for screening male sex hormone dependent/non-dependent hair growth-regulating agent for one kind, it is characterized in that:
In the presence of analyte, the hair follicle of flank, pars inguinalis, shoulder or the buttocks of pig is carried out organ culture, the material that promotes or suppress the growth of this hair follicle is estimated or selected.
Applications Claiming Priority (9)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2007042560 | 2007-02-22 | ||
| JP2007-042560 | 2007-02-22 | ||
| JP2007042560 | 2007-02-22 | ||
| JP2007295220 | 2007-11-14 | ||
| JP2007295220 | 2007-11-14 | ||
| JP2007-295220 | 2007-11-14 | ||
| JP2007-338801 | 2007-12-28 | ||
| JP2007338801 | 2007-12-28 | ||
| JP2007338801A JP4373468B2 (en) | 2007-02-22 | 2007-12-28 | Method for evaluating or screening hair growth regulator |
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| CN101251528A true CN101251528A (en) | 2008-08-27 |
| CN101251528B CN101251528B (en) | 2013-03-06 |
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| JP (1) | JP4373468B2 (en) |
| CN (1) | CN101251528B (en) |
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| JP6220506B2 (en) * | 2012-09-19 | 2017-10-25 | 花王株式会社 | Hair suppressant |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0843375A (en) * | 1994-08-03 | 1996-02-16 | Pola Chem Ind Inc | Hair tonic evaluating method |
| WO2003104443A2 (en) * | 2002-06-05 | 2003-12-18 | Rolf Hoffmann | Hair follicle mesenchymal stem cells and use thereof |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1062308C (en) * | 1997-12-09 | 2001-02-21 | 中国科学院新疆化学研究所 | Hair follicle culture establishing external drug screening and testing system |
| JP4254242B2 (en) * | 2003-01-08 | 2009-04-15 | 住友電気工業株式会社 | Hair growth activity evaluation method and hair growth activity evaluation kit |
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2007
- 2007-12-28 JP JP2007338801A patent/JP4373468B2/en not_active Expired - Fee Related
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2008
- 2008-02-18 CN CN 200810080430 patent/CN101251528B/en not_active Expired - Fee Related
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0843375A (en) * | 1994-08-03 | 1996-02-16 | Pola Chem Ind Inc | Hair tonic evaluating method |
| WO2003104443A2 (en) * | 2002-06-05 | 2003-12-18 | Rolf Hoffmann | Hair follicle mesenchymal stem cells and use thereof |
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| Publication number | Publication date |
|---|---|
| JP4373468B2 (en) | 2009-11-25 |
| JP2009139353A (en) | 2009-06-25 |
| DE602008002079D1 (en) | 2010-09-23 |
| CN101251528B (en) | 2013-03-06 |
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