Abstract
We succeeded in serially culturing cells derived from human sebaceous gland (sebocytes) under serum-free conditions. Sebaceous glands were isolated from dispase-treated facial skin specimens and cultured using two different methods, explant culture and dispersed cell culture, in KGM. In both types of culture the sebocytes proliferated rapidly without a biological feeder layer or specific matrices. It was possible to cultivate the cells serially for at least three passages (explant culture) and six passages (dispersed cell culture), and to store the cells in liquid nitrogen with good recovery. Analytical thin-layer chromatography revealed that the cells synthesized a large amount of sebum-specific lipids, squalene and wax esters in vitro. Both testosterone and 5α-dihydrotestosterone significantly stimulated the proliferation of the sebocytes. The methods presented here may permit a large series of studies on the function of sebaceous glands or cells derived from sebaceous glands.
Similar content being viewed by others
References
Akamatsu H, Zouboulis C, Orfanos C (1992) Control of human sebocyte proliferation in vitro by testosterone and 5-alpha-dihydrotestosterone is dependent on the localization of sebaceous glands. J Invest Dermatol 99:509–511
Arase S, Katoh S, Sadamoto Y, Nakanishi H, Urano Y, Fujie K, Takeda K (1991) Culture of human outer root sheath cells from plucked hair follicles in serum-free conditions. J Dermatol Sci 2:66–70
Bajor J, Siegel D, Kleinberg I, Simon M (1994) The in vitro proliferative potential of the human sebocyte (abstract). J Invest Dermatol 102:564
Boyce S, Ham R (1985) Cultivation, frozen storage, and clonal growth of normal human epidermal keratinocytes in serum-free media. J Tissue Culture Methods 9:83–93
Doran T, Baff R, Jacobs P, Pacia E (1991) Characterization of human sebaceous cells in vitro. J Invest Dermatol 96:341–348
Ishiyama M, Shiga M, Sasamoto K, Mizoguchi M, He P (1993) A new sulfonated tetrazolium salt that produces a highly watersoluble formazan dye. Chem Pharm Bull 41:1118–1122
Ishiyama M, Tominaga H, Shiga M, Sasamoto K, Ohkura Y, Ueno K (1995) Novel cell proliferation and cytotoxicity assays using a tetrazolium salt that produces a water-soluble formazan dye. In Vitro Toxicol 8:187–190
Johnson M, Karasek M (1987) Stimulation of lipid synthesis in human sebacytes by serum factors (abstract). J Invest Dermatol 88:496
Karasek M (1986) Growth characteristics of human sebaceous gland cells in cell culture (abstract). Clin Res 34:416A
Karasek M (1986) Isolation and characterization of cells from the human sebaceous gland (abstract). In Vitro Cell Dev Biol 22 (part II): 22A
Karasek M, Charlton M (1977) Isolation and growth for rabbit and human sebaceous gland cells in cell culture (abstract). J Invest Dermatol 68:234
Karasek M, Charlton M (1982) In vitro growth and serial cultivation of normal human sebaceous gland cells (abstract). Clin Res 30:263A
Laurent S, Mednieks M, Rosenfield R (1992) Growth of sebaceous cells in monolayer culture. In Vitro Cell Dev Biol 28A: 83–89
Nagae H, Nakanishi H, Urano Y, Arase S (1995) Serial cultivation of human nail matrix cells under serum-free conditions. J Dermatol 22:560–566
Roehm N, Rodgers G, Hatfield S, Glasebrook A (1991) An improved colorimetric assay for cell proliferation and viability utilizing the tetrazolium salt XTT. J Immunol Methods 142: 257–265
Uchida N, Oura H, Nakanishi H, Urano Y, Arase S (1993) Dispersed cell culture of human sweat duct cells under serum-free conditions. J Dermatol 20:684–690
Wille J, Pittelkow M, Shipley G, Scott R (1984) Integrated control of growth and differentiation of normal human prokeratinocytes cultured in serum-free medium: clonal analyses, growth kinetics and cell cycle studies. J Cell Physiol 121: 31–44
Xia L, Zouboulis C, Detmar M, Mayer-da-Silva A, Stadlar R, Orfanos C (1989) Isolation of human sebaceous glands and cultivation of sebaceous gland-derived cells as an in vitro model. J Invest Dermatol 93:315–321
Zouboulis C (1992) Sebaceous cells in monolayer culture (letter). In Vitro Cell Dev Biol 28A:699
Zouboulis C, Xia L, Detmar M, Bogdanoff B, Giannakopoulos G, Gollnick H, Orfanos CE (1991) Culture of human sebocytes and markers of sebocytic differentiation in vitro. Skin Pharmacol 4:74–83
Zouboulis C, Krieter A, Gollnick H, Mischke D, Orfanos C (1994) Progressive differentiation of human sebocytes in vitro is characterized by increasing cell size and altering antigen expression and is regulated by culture duration and retinoids. Exp Dermatol 3:151–160
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Fujie, T., Shikiji, T., Uchida, N. et al. Culture of cells derived from the human sebaceous gland under serum-free conditions without a biological feeder layer or specific matrices. Arch Dermatol Res 288, 703–708 (1996). https://doi.org/10.1007/BF02505281
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF02505281