Establishment of an Efficient Primary Culture System for Human Hair Follicle Stem Cells Using the Rho-Associated Protein Kinase Inhibitor Y-27632
BackgroundHair follicle tissue engineering is a promising strategy for treating hair loss. Human hair follicle stem cells (hHFSCs), which play a key role in the hair cycle, have potential applications in regenerative medicine. However, previous studies did not achieve efficient hHFSC expansion in vi...
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| Format: | Article |
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Frontiers Media S.A.
2021-03-01
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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.2021.632882/full |
| _version_ | 1819320439540809728 |
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| author | Lihong Wen Yong Miao Zhexiang Fan Jiarui Zhang Yixuan Guo Damao Dai Junfei Huang Zhen Liu Ruosi Chen Zhiqi Hu |
| author_facet | Lihong Wen Yong Miao Zhexiang Fan Jiarui Zhang Yixuan Guo Damao Dai Junfei Huang Zhen Liu Ruosi Chen Zhiqi Hu |
| author_sort | Lihong Wen |
| collection | DOAJ |
| description | BackgroundHair follicle tissue engineering is a promising strategy for treating hair loss. Human hair follicle stem cells (hHFSCs), which play a key role in the hair cycle, have potential applications in regenerative medicine. However, previous studies did not achieve efficient hHFSC expansion in vitro using feeder cells. Therefore, there is a need to develop an efficient primary culture system for the expansion and maintenance of hHFSCs.MethodsThe hHFSCs were obtained by two-step proteolytic digestion combined with microscopy. The cell culture dishes were coated with human fibronectin and inoculated with hHFSCs. The hHFSCs were harvested using a differential enrichment procedure. The effect of Rho-associated protein kinase (ROCK) inhibitor Y-27632, supplemented in keratinocyte serum-free medium (K-SFM), on adhesion, proliferation, and stemness of hHFSCs and the underlying molecular mechanisms were evaluated.ResultsThe hHFSCs cultured in K-SFM, supplemented with Y-27632, exhibited enhanced adhesion and proliferation. Additionally, Y-27632 treatment maintained the stemness of hHFSCs and promoted the ability of hHFSCs to regenerate hair follicles in vivo. However, Y-27632-induced proliferation and stemness in hHFSCs were conditional and reversible. Furthermore, Y-27632 maintained propagation and stemness of hHFSCs through the ERK/MAPK pathway.ConclusionAn efficient short-term culture system for primary hHFSCs was successfully established using human fibronectin and the ROCK inhibitor Y-27632, which promoted the proliferation, maintained the stemness of hHFSCs and promoted the ability to regenerate hair follicles in vivo. The xenofree culturing method used in this study provided a large number of high-quality seed cells, which have applications in hair follicle tissue engineering and stem cell therapy. |
| first_indexed | 2024-12-24T11:19:36Z |
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| institution | Directory Open Access Journal |
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| language | English |
| last_indexed | 2024-12-24T11:19:36Z |
| publishDate | 2021-03-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cell and Developmental Biology |
| spelling | doaj.art-084aa353ee1a40fb85b7ef94babd7e692022-12-21T16:58:17ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2021-03-01910.3389/fcell.2021.632882632882Establishment of an Efficient Primary Culture System for Human Hair Follicle Stem Cells Using the Rho-Associated Protein Kinase Inhibitor Y-27632Lihong Wen0Yong Miao1Zhexiang Fan2Jiarui Zhang3Yixuan Guo4Damao Dai5Junfei Huang6Zhen Liu7Ruosi Chen8Zhiqi Hu9Department of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, ChinaDepartment of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, ChinaDepartment of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, ChinaDepartment of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, ChinaDepartment of Plastic and Burn Surgery, Shenzhen Hospital, Southern Medical University, Shenzhen, ChinaDepartment of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, ChinaDepartment of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, ChinaDepartment of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, ChinaDepartment of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, ChinaDepartment of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, ChinaBackgroundHair follicle tissue engineering is a promising strategy for treating hair loss. Human hair follicle stem cells (hHFSCs), which play a key role in the hair cycle, have potential applications in regenerative medicine. However, previous studies did not achieve efficient hHFSC expansion in vitro using feeder cells. Therefore, there is a need to develop an efficient primary culture system for the expansion and maintenance of hHFSCs.MethodsThe hHFSCs were obtained by two-step proteolytic digestion combined with microscopy. The cell culture dishes were coated with human fibronectin and inoculated with hHFSCs. The hHFSCs were harvested using a differential enrichment procedure. The effect of Rho-associated protein kinase (ROCK) inhibitor Y-27632, supplemented in keratinocyte serum-free medium (K-SFM), on adhesion, proliferation, and stemness of hHFSCs and the underlying molecular mechanisms were evaluated.ResultsThe hHFSCs cultured in K-SFM, supplemented with Y-27632, exhibited enhanced adhesion and proliferation. Additionally, Y-27632 treatment maintained the stemness of hHFSCs and promoted the ability of hHFSCs to regenerate hair follicles in vivo. However, Y-27632-induced proliferation and stemness in hHFSCs were conditional and reversible. Furthermore, Y-27632 maintained propagation and stemness of hHFSCs through the ERK/MAPK pathway.ConclusionAn efficient short-term culture system for primary hHFSCs was successfully established using human fibronectin and the ROCK inhibitor Y-27632, which promoted the proliferation, maintained the stemness of hHFSCs and promoted the ability to regenerate hair follicles in vivo. The xenofree culturing method used in this study provided a large number of high-quality seed cells, which have applications in hair follicle tissue engineering and stem cell therapy.https://www.frontiersin.org/articles/10.3389/fcell.2021.632882/fullhuman hair follicle stem cellshair follicle tissue engineeringY-27632primary culture systemERK/MAPK pathway |
| spellingShingle | Lihong Wen Yong Miao Zhexiang Fan Jiarui Zhang Yixuan Guo Damao Dai Junfei Huang Zhen Liu Ruosi Chen Zhiqi Hu Establishment of an Efficient Primary Culture System for Human Hair Follicle Stem Cells Using the Rho-Associated Protein Kinase Inhibitor Y-27632 Frontiers in Cell and Developmental Biology human hair follicle stem cells hair follicle tissue engineering Y-27632 primary culture system ERK/MAPK pathway |
| title | Establishment of an Efficient Primary Culture System for Human Hair Follicle Stem Cells Using the Rho-Associated Protein Kinase Inhibitor Y-27632 |
| title_full | Establishment of an Efficient Primary Culture System for Human Hair Follicle Stem Cells Using the Rho-Associated Protein Kinase Inhibitor Y-27632 |
| title_fullStr | Establishment of an Efficient Primary Culture System for Human Hair Follicle Stem Cells Using the Rho-Associated Protein Kinase Inhibitor Y-27632 |
| title_full_unstemmed | Establishment of an Efficient Primary Culture System for Human Hair Follicle Stem Cells Using the Rho-Associated Protein Kinase Inhibitor Y-27632 |
| title_short | Establishment of an Efficient Primary Culture System for Human Hair Follicle Stem Cells Using the Rho-Associated Protein Kinase Inhibitor Y-27632 |
| title_sort | establishment of an efficient primary culture system for human hair follicle stem cells using the rho associated protein kinase inhibitor y 27632 |
| topic | human hair follicle stem cells hair follicle tissue engineering Y-27632 primary culture system ERK/MAPK pathway |
| url | https://www.frontiersin.org/articles/10.3389/fcell.2021.632882/full |
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