Revisiting the Experimental Methods for Human Skin T-Cell Analysis
Tissue-resident memory T cells exist in both the epidermis and the dermis in human skin. To analyze these cells, the skin needs to be incubated with dispase II to separate the two layers, that is, the epidermis and the dermis. The next step varies among researchers; the subsequent enzymatic digestio...
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2022-07-01
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| Series: | JID Innovations |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2667026722000339 |
| _version_ | 1828189162262495232 |
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| author | Takuya Sato Youichi Ogawa Aoha Ishikawa Yuka Nagasaka Manao Kinoshita Ichiro Shiokawa Shinji Shimada Akira Momosawa Tatsuyoshi Kawamura |
| author_facet | Takuya Sato Youichi Ogawa Aoha Ishikawa Yuka Nagasaka Manao Kinoshita Ichiro Shiokawa Shinji Shimada Akira Momosawa Tatsuyoshi Kawamura |
| author_sort | Takuya Sato |
| collection | DOAJ |
| description | Tissue-resident memory T cells exist in both the epidermis and the dermis in human skin. To analyze these cells, the skin needs to be incubated with dispase II to separate the two layers, that is, the epidermis and the dermis. The next step varies among researchers; the subsequent enzymatic digestion of the two layers is popular, whereas the spontaneous migration method can also be done. Scraping of these layers to yield skin T cells may reduce antigen modulation. This study aimed to determine each method’s limitations. Dispase II incubation itself cleaves T-cell antigens. Therefore, further enzymatic digestion with collagenases strongly cleaves antigens. The scraping method yields skin T cells that are affected by dispase II as it is. However, skin T-cell yield is low. The spontaneous migration method recovers and/or upregulates antigens with T-cell activation and loses ∼20% of T cells in the floating sheets. However, there was no prominent bias regarding CD103 expression between emigrants and the remaining T cells in the sheets. There were 104 and 105 CD3+ T cells per 1 cm2 of the epidermis and upper dermis, respectively. Collectively, each method has strengths and limitations to analyze both the epidermal and dermal T cells. |
| first_indexed | 2024-04-12T08:01:12Z |
| format | Article |
| id | doaj.art-9313daa348894af584c7a0451817dd24 |
| institution | Directory Open Access Journal |
| issn | 2667-0267 |
| language | English |
| last_indexed | 2024-04-12T08:01:12Z |
| publishDate | 2022-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | JID Innovations |
| spelling | doaj.art-9313daa348894af584c7a0451817dd242022-12-22T03:41:19ZengElsevierJID Innovations2667-02672022-07-0124100125Revisiting the Experimental Methods for Human Skin T-Cell AnalysisTakuya Sato0Youichi Ogawa1Aoha Ishikawa2Yuka Nagasaka3Manao Kinoshita4Ichiro Shiokawa5Shinji Shimada6Akira Momosawa7Tatsuyoshi Kawamura8Department of Dermatology, Faculty of Medicine, University of Yamanashi, Yamanashi, JapanDepartment of Dermatology, Faculty of Medicine, University of Yamanashi, Yamanashi, Japan; Correspondence: Youichi Ogawa, Department of Dermatology, Faculty of Medicine, University of Yamanashi, 1110 Shimokato, Chuo, Yamanashi 409-3898, JapanDepartment of Plastic Surgery, Faculty of Medicine, University of Yamanashi, Yamanashi, JapanDepartment of Plastic Surgery, Faculty of Medicine, University of Yamanashi, Yamanashi, JapanDepartment of Dermatology, Faculty of Medicine, University of Yamanashi, Yamanashi, JapanDepartment of Plastic Surgery, Faculty of Medicine, University of Yamanashi, Yamanashi, JapanDepartment of Dermatology, Faculty of Medicine, University of Yamanashi, Yamanashi, JapanDepartment of Plastic Surgery, Faculty of Medicine, University of Yamanashi, Yamanashi, JapanDepartment of Dermatology, Faculty of Medicine, University of Yamanashi, Yamanashi, JapanTissue-resident memory T cells exist in both the epidermis and the dermis in human skin. To analyze these cells, the skin needs to be incubated with dispase II to separate the two layers, that is, the epidermis and the dermis. The next step varies among researchers; the subsequent enzymatic digestion of the two layers is popular, whereas the spontaneous migration method can also be done. Scraping of these layers to yield skin T cells may reduce antigen modulation. This study aimed to determine each method’s limitations. Dispase II incubation itself cleaves T-cell antigens. Therefore, further enzymatic digestion with collagenases strongly cleaves antigens. The scraping method yields skin T cells that are affected by dispase II as it is. However, skin T-cell yield is low. The spontaneous migration method recovers and/or upregulates antigens with T-cell activation and loses ∼20% of T cells in the floating sheets. However, there was no prominent bias regarding CD103 expression between emigrants and the remaining T cells in the sheets. There were 104 and 105 CD3+ T cells per 1 cm2 of the epidermis and upper dermis, respectively. Collectively, each method has strengths and limitations to analyze both the epidermal and dermal T cells.http://www.sciencedirect.com/science/article/pii/S2667026722000339 |
| spellingShingle | Takuya Sato Youichi Ogawa Aoha Ishikawa Yuka Nagasaka Manao Kinoshita Ichiro Shiokawa Shinji Shimada Akira Momosawa Tatsuyoshi Kawamura Revisiting the Experimental Methods for Human Skin T-Cell Analysis JID Innovations |
| title | Revisiting the Experimental Methods for Human Skin T-Cell Analysis |
| title_full | Revisiting the Experimental Methods for Human Skin T-Cell Analysis |
| title_fullStr | Revisiting the Experimental Methods for Human Skin T-Cell Analysis |
| title_full_unstemmed | Revisiting the Experimental Methods for Human Skin T-Cell Analysis |
| title_short | Revisiting the Experimental Methods for Human Skin T-Cell Analysis |
| title_sort | revisiting the experimental methods for human skin t cell analysis |
| url | http://www.sciencedirect.com/science/article/pii/S2667026722000339 |
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