A 2D and 3D melanogenesis model with human primary cells induced by tyrosine

Authors

  • Maryana S. Branquinho University of Sao Paulo
  • Maysa B. Silva University of Sao Paulo
  • Jacqueline C. Silva University of Sao Paulo
  • Maria C. Sales University of Sao Paulo
  • Silvia B. Barros University of Sao Paulo
  • Silvya S. Maria-Engler University of Sao Paulo
  • Ana Campa University of Sao Paulo

DOI:

https://doi.org/10.14440/jbm.2020.327

Keywords:

co-culture, keratinocyte, melanin, melanocyte, pigmentation

Abstract

Research on melanogenesis, its regulation in health and disease, and the discovery of new molecules with pigmenting and depigmenting activities use different models. Here we standardize a protocol based on previous ones using primary human melanocytes and keratinocytes in co-cultures, in which melanogenesis was induced under mild conditions by the addition of tyrosine plus ammonium chloride (NH4Cl). The expression of MITF, TYR, TYRP1, and Melan-A as well as melanin content were measured. Furthermore, we extended this study to a reconstructed 3D model. Pigmentation was visually observable and melanosomes were identified by Fontana-Masson staining by the addition of tyrosine plus NH4Cl during the stratification phase. The 2D and 3D protocols proposed here circumvent limitations of previous models, using human primary cells and mild conditions for melanogenesis. These protocols offer a viable, robust, simple, and animal-free investigational option for human skin pigmentation studies and screening tests for new compounds that modulate pigmentation.

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Published

2020-07-06

How to Cite

1.
Branquinho MS, Silva MB, Silva JC, Sales MC, Barros SB, Maria-Engler SS, Campa A. A 2D and 3D melanogenesis model with human primary cells induced by tyrosine. J Biol Methods [Internet]. 2020Jul.6 [cited 2021Aug.2];7(3):e134. Available from: https://jbmethods.org/jbm/article/view/327

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