“Two-Cell Assemblage” Assay: A Simple in vitro Method for Screening Hair Growth-Promoting Compounds
Alopecia arises due to inadequate hair follicle (HF) stem cell activation or proliferation, resulting in prolongation of the telogen phase of the hair cycle. Increasing therapeutic and cosmetic demand for alleviating alopecia has driven research toward the discovery or synthesis of novel compounds t...
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| Format: | Article |
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Frontiers Media S.A.
2020-11-01
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| Series: | Frontiers in Cell and Developmental Biology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcell.2020.581528/full |
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| author | Sunhyae Jang Sunhyae Jang Sunhyae Jang Jungyoon Ohn Jungyoon Ohn Jungyoon Ohn Bo Mi Kang Bo Mi Kang Bo Mi Kang Minji Park Minji Park Minji Park Kyu Han Kim Kyu Han Kim Kyu Han Kim Ohsang Kwon Ohsang Kwon Ohsang Kwon |
| author_facet | Sunhyae Jang Sunhyae Jang Sunhyae Jang Jungyoon Ohn Jungyoon Ohn Jungyoon Ohn Bo Mi Kang Bo Mi Kang Bo Mi Kang Minji Park Minji Park Minji Park Kyu Han Kim Kyu Han Kim Kyu Han Kim Ohsang Kwon Ohsang Kwon Ohsang Kwon |
| author_sort | Sunhyae Jang |
| collection | DOAJ |
| description | Alopecia arises due to inadequate hair follicle (HF) stem cell activation or proliferation, resulting in prolongation of the telogen phase of the hair cycle. Increasing therapeutic and cosmetic demand for alleviating alopecia has driven research toward the discovery or synthesis of novel compounds that can promote hair growth by inducing HF stem cell activation or proliferation and initiating the anagen phase. Although several methods for evaluating the hair growth-promoting effects of candidate compounds are being used, most of these methods are difficult to use for large scale simultaneous screening of various compounds. Herein, we introduce a simple and reliable in vitro assay for the simultaneous screening of the hair growth-promoting effects of candidate compounds on a large scale. In this study, we first established a 3D co-culture system of human dermal papilla (hDP) cells and human outer root sheath (hORS) cells in an ultra-low attachment 96-well plate, where the two cell types constituted a polar elongated structure, named “two-cell assemblage (TCA).” We observed that the long axis length of the TCA gradually increased for 5 days, maintaining biological functional integrity as reflected by the increased expression levels of hair growth-associated genes after treatment with hair growth-promoting molecules. Interestingly, the elongation of the TCA was more prominent following treatment with the hair growth-promoting molecules (which occurred in a dose-dependent manner), compared to the control group (p < 0.05). Accordingly, we set the long axis length of the TCA as an endpoint of this assay, using a micro confocal high-content imaging system to measure the length, which can provide reproducible and reliable results in an adequate timescale. The advantages of this assay are: (i) it is physiologically and practically advantageous as it uses 3D cultured two-type human cells which are easily available; (ii) it is simple as it uses length as the only endpoint; and (iii) it is a high throughput system, which screens various compounds simultaneously. In conclusion, the “TCA” assay could serve as an easy and reliable method to validate the hair growth-promoting effect of a large volume of library molecules. |
| first_indexed | 2024-12-17T03:29:43Z |
| format | Article |
| id | doaj.art-c33ec20bf6094918a1382d3a4e2aa5ab |
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| issn | 2296-634X |
| language | English |
| last_indexed | 2024-12-17T03:29:43Z |
| publishDate | 2020-11-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cell and Developmental Biology |
| spelling | doaj.art-c33ec20bf6094918a1382d3a4e2aa5ab2022-12-21T22:05:18ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2020-11-01810.3389/fcell.2020.581528581528“Two-Cell Assemblage” Assay: A Simple in vitro Method for Screening Hair Growth-Promoting CompoundsSunhyae Jang0Sunhyae Jang1Sunhyae Jang2Jungyoon Ohn3Jungyoon Ohn4Jungyoon Ohn5Bo Mi Kang6Bo Mi Kang7Bo Mi Kang8Minji Park9Minji Park10Minji Park11Kyu Han Kim12Kyu Han Kim13Kyu Han Kim14Ohsang Kwon15Ohsang Kwon16Ohsang Kwon17Laboratory of Cutaneous Aging and Hair Research, Clinical Research Institute, Seoul National University Hospital, Seoul, South KoreaInstitute of Human Environment Interface Biology, Seoul National University, Seoul, South KoreaDepartment of Dermatology, Seoul National University College of Medicine, Seoul, South KoreaLaboratory of Cutaneous Aging and Hair Research, Clinical Research Institute, Seoul National University Hospital, Seoul, South KoreaInstitute of Human Environment Interface Biology, Seoul National University, Seoul, South KoreaDepartment of Dermatology, Seoul National University College of Medicine, Seoul, South KoreaLaboratory of Cutaneous Aging and Hair Research, Clinical Research Institute, Seoul National University Hospital, Seoul, South KoreaInstitute of Human Environment Interface Biology, Seoul National University, Seoul, South KoreaDepartment of Dermatology, Seoul National University College of Medicine, Seoul, South KoreaLaboratory of Cutaneous Aging and Hair Research, Clinical Research Institute, Seoul National University Hospital, Seoul, South KoreaInstitute of Human Environment Interface Biology, Seoul National University, Seoul, South KoreaDepartment of Dermatology, Seoul National University College of Medicine, Seoul, South KoreaLaboratory of Cutaneous Aging and Hair Research, Clinical Research Institute, Seoul National University Hospital, Seoul, South KoreaInstitute of Human Environment Interface Biology, Seoul National University, Seoul, South KoreaDepartment of Dermatology, Seoul National University College of Medicine, Seoul, South KoreaLaboratory of Cutaneous Aging and Hair Research, Clinical Research Institute, Seoul National University Hospital, Seoul, South KoreaInstitute of Human Environment Interface Biology, Seoul National University, Seoul, South KoreaDepartment of Dermatology, Seoul National University College of Medicine, Seoul, South KoreaAlopecia arises due to inadequate hair follicle (HF) stem cell activation or proliferation, resulting in prolongation of the telogen phase of the hair cycle. Increasing therapeutic and cosmetic demand for alleviating alopecia has driven research toward the discovery or synthesis of novel compounds that can promote hair growth by inducing HF stem cell activation or proliferation and initiating the anagen phase. Although several methods for evaluating the hair growth-promoting effects of candidate compounds are being used, most of these methods are difficult to use for large scale simultaneous screening of various compounds. Herein, we introduce a simple and reliable in vitro assay for the simultaneous screening of the hair growth-promoting effects of candidate compounds on a large scale. In this study, we first established a 3D co-culture system of human dermal papilla (hDP) cells and human outer root sheath (hORS) cells in an ultra-low attachment 96-well plate, where the two cell types constituted a polar elongated structure, named “two-cell assemblage (TCA).” We observed that the long axis length of the TCA gradually increased for 5 days, maintaining biological functional integrity as reflected by the increased expression levels of hair growth-associated genes after treatment with hair growth-promoting molecules. Interestingly, the elongation of the TCA was more prominent following treatment with the hair growth-promoting molecules (which occurred in a dose-dependent manner), compared to the control group (p < 0.05). Accordingly, we set the long axis length of the TCA as an endpoint of this assay, using a micro confocal high-content imaging system to measure the length, which can provide reproducible and reliable results in an adequate timescale. The advantages of this assay are: (i) it is physiologically and practically advantageous as it uses 3D cultured two-type human cells which are easily available; (ii) it is simple as it uses length as the only endpoint; and (iii) it is a high throughput system, which screens various compounds simultaneously. In conclusion, the “TCA” assay could serve as an easy and reliable method to validate the hair growth-promoting effect of a large volume of library molecules.https://www.frontiersin.org/articles/10.3389/fcell.2020.581528/fullalopeciahair folliclein vitro assayouter root sheath cellscreening assaydermal papilla cell |
| spellingShingle | Sunhyae Jang Sunhyae Jang Sunhyae Jang Jungyoon Ohn Jungyoon Ohn Jungyoon Ohn Bo Mi Kang Bo Mi Kang Bo Mi Kang Minji Park Minji Park Minji Park Kyu Han Kim Kyu Han Kim Kyu Han Kim Ohsang Kwon Ohsang Kwon Ohsang Kwon “Two-Cell Assemblage” Assay: A Simple in vitro Method for Screening Hair Growth-Promoting Compounds Frontiers in Cell and Developmental Biology alopecia hair follicle in vitro assay outer root sheath cell screening assay dermal papilla cell |
| title | “Two-Cell Assemblage” Assay: A Simple in vitro Method for Screening Hair Growth-Promoting Compounds |
| title_full | “Two-Cell Assemblage” Assay: A Simple in vitro Method for Screening Hair Growth-Promoting Compounds |
| title_fullStr | “Two-Cell Assemblage” Assay: A Simple in vitro Method for Screening Hair Growth-Promoting Compounds |
| title_full_unstemmed | “Two-Cell Assemblage” Assay: A Simple in vitro Method for Screening Hair Growth-Promoting Compounds |
| title_short | “Two-Cell Assemblage” Assay: A Simple in vitro Method for Screening Hair Growth-Promoting Compounds |
| title_sort | two cell assemblage assay a simple in vitro method for screening hair growth promoting compounds |
| topic | alopecia hair follicle in vitro assay outer root sheath cell screening assay dermal papilla cell |
| url | https://www.frontiersin.org/articles/10.3389/fcell.2020.581528/full |
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