Isolation, culture and phenotypic characterization of human sweat gland epithelial cells

Gao,Yunhe; Li,Meiying; Zhang,Xueyan; Bai,Tingting; Chi,Guanfan; Liu,Jin  Yu; Li,Yulin

doi:10.3892/ijmm.2014.1851

October 2014 Volume 34 Issue 4

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October 2014 Volume 34 Issue 4

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Isolation, culture and phenotypic characterization of human sweat gland epithelial cells

Authors:
- Yunhe Gao
- Meiying Li
- Xueyan Zhang
- Tingting Bai
- Guanfan Chi
- Jin Yu Liu
- Yulin Li
View Affiliations / Copyright

Affiliations: The Key Laboratory of Pathobiology, Ministry of Education, College of Basic Medical Sciences, Jilin University, Changchun, Jilin 130021, P.R. China

Copyright: © Gao et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].
Pages: 997-1003
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Published online on: July 14, 2014

https://doi.org/10.3892/ijmm.2014.1851
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Abstract

Sweat gland epithelial cells (SGECs) have been identified as essential for the regeneration of sweat glands and for the construction of skin substitutes containing skin appendages. Consequently, the isolation, culture and phenotypic characterization of SGECs are of paramount importance. In the present study study, human sweat glands were isolated by pipetting under a phase contrast microscope following digestion with collagenase type Ⅰ. Subsequently, a microscopic organ culture technique was used for the primary culture of human SGECs, and the culture conditions were modified in order to achieve optimal cell growth status. Primary SGECs were identified based on their expression of markers specific for sweat glands, including carcinoembryonic antigen (CEA), CK7, CK8, CK14, CK15, CK18 and CK19. We explored the possible presence of stem cells in human sweat glands by detecting their expression of leucine‑rich repeat-containing G protein‑coupled receptor 5 (LGR5). Primary SGECs achieved a good growth state when cultured under serum‑free conditions. After one passage, the cells cultured in keratinocyte serum-free medium with 1% fetal bovine serum (FBS) still showed a prominent proliferative activity. Phenotypic analysis by immunofluorescence microscopy, reverse transcription-polymerase chain reaction (RT‑PCR), and western blot analysis demonstrated the expression of sweat gland‑specific markers, including CEA, CK7, CK8, CK14, CK15, CK18 and CK19. In addition, RT‑PCR and immunochemistry detected the expression of LGR5. In comparison with traditional serum‑containing conditions, serum‑free culture provides the preferred culture conditions for human SGECs. LGR5 is a novel marker that identifies human sweat gland‑derived stem cells.

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Journals

International Journal of Molecular Medicine

International Journal of Oncology

Molecular Medicine Reports

Oncology Reports

Experimental and Therapeutic Medicine

Oncology Letters

Biomedical Reports

Molecular and Clinical Oncology

World Academy of Sciences Journal

International Journal of Functional Nutrition

International Journal of Epigenetics

Medicine International

Isolation, culture and phenotypic characterization of human sweat gland epithelial cells

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