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Isolation, culture, and verification of human sweat gland epithelial cells

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Abstract

Human sweat gland epithelial cells (SGECs) have been isolated and grown in vitro, However, slow proliferation makes the culture of these cells extremely difficult. The present study was carried out to explore the modified culture medium for SGECs in vitro. Full-thickness skin samples were minced (1 mm3) and digested overnight with type II collagenase. The gland coils were removed under an inverted phase-contrast microscope. An adherent culture method was used to isolate and culture SGECs. Staining with hematoxylin and eosin was performed, followed by observation of the morphologic features of these cells. Immunofluorescence staining with antibodies to cytokeratins CK7, CK18, and CK19 and carcinoembryonic antigen (CEA) was performed to verify the presence of SGECs. Growth curves by MTT were created for cells grown in serum-free keratinocyte medium and in modified keratinocyte medium containing 2.5% fetal bovine serum (FBS). One week after culturing, the cells grew well and were polygonal or irregular in shape by inverted phase contrast microscopy. Cell fusion, with a characteristic paving-stone arrangement, reached 100% after approximately 3 weeks in culture. Immunofluorescence staining indicated expression of CK7, CK18, CK19, and CEA. Compared with SGECs grown in serum-free keratinocyte medium, the proliferation of SGECs grown in modified culture medium with low concentration of FBS at days 6, 9, and 12 was significantly accelerated (p < 0.05). This study suggests that keratinocyte medium supplemented with 2.5% FBS is effective and suitable for the culture of SGECs.

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Acknowledgments

The technical assistance of Dr. Yin Zhou is gratefully acknowledged. The study was supported by grants from the National health public welfare special scientific research foundation of China (200802066) and the People’s Liberation Army health science research foundation of China (06Z054).

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Correspondence to Jiake Chai.

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Ran Tao and Yanfu Han contributed equally to this work.

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Tao, R., Han, Y., Chai, J. et al. Isolation, culture, and verification of human sweat gland epithelial cells. Cytotechnology 62, 489–495 (2010). https://doi.org/10.1007/s10616-010-9303-z

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  • DOI: https://doi.org/10.1007/s10616-010-9303-z

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