In Vitro Evaluation Method to Monitor Pathogenic Acne Vulgaris by Culturing Keratinocytes with Sebocytes

Park, Mi Kyung; Park, Kui Young; Kim, Beom Joon; Kim, Myeung Nam; Kim, Jeong Hwan; Koh, Hyun Ju; Park, Won Seok

Abstract

BACKGROUND
Acne vulgaris is a chronic inflammatory disease of the pilosebaceous unit. Keratinocytes and sebocytes are the two major cell types involved in the induction of acne within the pilosebaceous unit. Thus, keratinocytes or sebocytes cultured alone may not accurately reflect the physiology of the acne lesion.
OBJECTIVE
The aim of this study was to examine inflammatory cytokine production and neutral lipid content in keratinocyte and sebocyte co-cultures treated with acne etiologic stimulants.
METHODS
Spontaneously immortalized, non-tumorigenic human HaCaT keratinocytes and immortalized human facial SZ95 sebocytes were seeded in 24-well culture plates (Nunc, Wiesbaden, Germany). Secretion of inflammatory cytokines by each cell type was analyzed by enzyme-linked immunosorbent assay, and neutral lipid production in the cells was measured by Oil Red O staining. Cells were then simultaneously treated with multiple stimulants at various concentrations, and triglyceride and interleukin 8 (IL-8) levels were measured.
RESULTS
Arachidonic acid, linoleic acid, dihydrotestosterone and P. acnes extract all induced IL-8 production. De novo synthesis of sebaceous lipids was observed more frequently in the keratinocyte/sebocyte co-culture than that in the sebocyte monoculture. Higher levels of IL-8 were detected in the multi-stimulant system than those in the mono stimulant system.
CONCLUSION
The results suggest that co-culture of keratinocytes and sebocytes is more representative of the clinical condition associated with the development of acne lesions than culture of either cell type alone. The co-culture system used in this study is applicable to research acne vulgaris pathogenesis and to screen active drugs for treating this condition.