Culturing of kerationocytes in serum-free medium

• Main Authors: Mohammad Shokerzadeh, Mahnaz Mahmoudi Rad, Mansoureh Mirzaei, Nariman Mosef3, Maedeh Ahangari
• Format: Article
• Language: English
• Published: Mazandaran University of Medical Sciences 2009-01-01
• Series: Journal of Mazandaran University of Medical Sciences
• Subjects: Keratinocyte; epidermal explants; serum-free medium
• Online Access: http://www.mazums.ac.ir/index.php?digital_library&do=downloadPdf&sid=757

(Received 9 February, 2009 ; Accepted 6 May, 2009)AbstractBackground and purpose: To establish a simple method for preparation of pure epidermal cell in serum-free medium, without co culturing with lethally irradiated 3T3 cells.Materials and methods: A piece of skin biopsy was taken from a rat. After trypsinazation to separate epidermal layer, the epidermis was cut into explants 1×1 mm and were laid at 50 mL on tissue cultured flasks. Then, the explants were covered with serum-free keratinocyte growth medium, while the culture medium was changed every two days. The keratinocyte colonies were expanded in flasks and proliferated to 70% confluency on day 21. After two subcultures, the cells were frozen.Results: Pure epidermal cells are established by this technique in a serum-free keratinocyte growth medium, under feeder cell-free condition.Conclusion: Culturing of keratinocytes in serum-free medium is proved to be a useful and simple method for keratinocyte isolation. The successful culturing of keratinocytes from a small skin biopsy can be useful in the treatment of major burn wounds.J Mazand Univ Med Sci 2009; 19(70): 67-71 (Persian)