Kubilus, J., Neal, P., Kaluzhny, Y., Sur, G., and Klausner, M. MatTek Corporation, Ashland, MA.

Human skin reconstructs are three dimensional in vitro models consisting of a co-culture of epidermal keratinocytes and melanocytes plated onto a dermal substrate. In contrast to cells in monolayer culture, tissue-engineered skin equivalents contain well-developed epidermal layers with normal melanocytes being distributed at the basement membrane that separates the epidermis from the dermis. The incidence of cutaneous melanoma (CM) has increased dramatically in the last few decades. Development of specific CM stage models (e.g. radial (RGP) or vertical (VGP) growth phases) with their specific microenvironments and cell-matrix and cell-cell communication are crucial for understanding changes leading to the metastatic CM phenotype and for the development of CM therapies. A serum-free (SF) culture system, in which the precise effects of growth factors and chemo-therapeutic compounds can be tested without interference from the multitude of undetermined components in serum, is presented. Single cell suspensions of normal human epidermal keratinocytes and melanoma cells from different stages of melanoma progression are seeded on acellular or fibroblast-contracted collagen gels and induced to differentiate in SF medium. Incorporation of the metastatic melanoma cell lines, A375 and SK-MEL-28, into the epidermal/acellular dermal cultures results in progressively growing clusters of melanoma cells in the basal/spinous layers. Similarly, when A375 cells are added to MatTek’s full thickness skin model (FTSM), nodes of melanoma cells develop in the epidermis by day 10. These clusters begin to invade the dermis by day 14. Incorporation of SK-MEL-28 into FTSM shows progressive disruption of the basement membrane. Finally, the Wistar metastatic cells (WM852) yielded a tumor invading the dermis on day 16. The development of three dimensional, serum-free tissue culture models, containing melanocytes at different stages of CM malignancy, will provide researchers with valuable tools to study, understand, and develop preventative and therapeutic treatments for this most serious cutaneous malignancy.


A375, Cutaneous melanoma (CM), EpiDerm-FT, EpiDermFT, Fetal bovine serum (FBS), Fibroblast (FB), Full thickness skin model (FTSM), Human metastatic melanoma (MM), In vitro, Keratinocytes, MelanoDerm, Melanocytes, Melanoma, NHEK, Normal human keratinocytes (NHK), Normal human melanocytes (NHM), Radial growth phase (RGP), SK-MEL-28, Serum-free (SF), Vertical growth phase (VGP), WM852, Wistar metastatic cells (WM852)

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