title: Development of melanocye-keratinocyte co-culture model for controls and vitiligo to assess regulators of pigmentation and melanocytes.
abstract: BACKGROUND
  There is a need to develop an in vitro skin models which can be used as alternative system for research and testing pharmacological products in place of laboratory animals. Therefore to study the biology and pathophysiology of pigmentation and vitiligo, reliable in vitro skin pigmentation models are required.
  
  
  OBJECTIVE
  In this study, we used primary cultured melanocytes and keratinocytes to prepare the skin co-culture model in control and vitiligo patients.
  
  
  METHODS
  The skin grafts were taken from control and patients of vitiligo. In vitro co-culture was prepared after culturing primary melanocytes and keratinocytes. Co- cultures were treated with melanogenic stimulators and inhibitors and after that tyrosinase assay, MTT assay and melanin content assay were performed.
  
  
  RESULTS
  Melanocytes and keratinocytes were successfully cultured from control and vitiligo patients and after that co-culture models were prepared. After treatment of co-culture model with melanogenic stimulator we found that tyrosinase activity, cell proliferation and melanin content increased whereas after treatment with melanogenic inhibitor, tyrosinase activity, cell proliferation and melanin content decreased. We also found some differences in the control co-culture model and vitiligo co-culture model.
  
  
  CONCLUSIONS
  We successfully constructed in vitro co-culture pigmentation model for control and vitiligo patients using primary cultured melanocytes and keratinocytes. The use of primary melanocytes and keratinocytes is more appropriate over the use of transformed cells. The only limitation of these models is that these can be used for screening small numbers of compounds.
authors.affiliation: Department of Dermatology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
authors.forename: Ravinder
authors.initials: R
authors.lastname: Kumar
authors.forename: Davinder
authors.initials: D
authors.lastname: Parsad
authors.forename: Amrinderjit
authors.initials: A
authors.lastname: Kanwar
authors.forename: Deepak
authors.initials: D
authors.lastname: Kaul
mesh_terms: D000704:Analysis of Variance; D049109:Cell Proliferation / Q000187:drug effects; D018920:Coculture Techniques; D006801:Humans; D006873:Hydroquinones / Q000494:pharmacology; D015603:Keratinocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D000077338:Latanoprost; D008543:Melanins / Q000819:agonists / Q000037:antagonists & inhibitors / Q000378:metabolism; D008544:Melanocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D008954:Models, Biological; D014442:Monophenol Monooxygenase / Q000187:drug effects / Q000378:metabolism; D010858:Pigmentation / Q000502:physiology; D011461:Prostaglandins F, Synthetic / Q000494:pharmacology; D011753:Pyrones / Q000494:pharmacology; D019089:Stem Cell Factor / Q000494:pharmacology; D014820:Vitiligo / Q000503:physiopathology*
chemical_list: D006873:Hydroquinones; D008543:Melanins; D011461:Prostaglandins F, Synthetic; D011753:Pyrones; D019089:Stem Cell Factor; C011890:kojic acid; D000077338:Latanoprost; D014442:Monophenol Monooxygenase; C031927:hydroquinone