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Comparison of two cellular harvesting methods for primary human oral culture of keratinocytes
Authors:Mª Fátima Guarizo Klingbeil  Marisa Roma Herson  Elier Broche Cristo  Décio dos Santos Pinto Jr  Daniele Yoshito  Monica Beatriz Mathor
Affiliation:1. Radiation Technology Center (CTR), Nuclear and Energetic Research Institute-IPEN/CNEN, Av. Lineu Prestes 2242, Cidade Universitária, S?o Paulo, Brazil
2. Medical School, Micro and Plastic Surgery Department, S?o Paulo University, Av. Dr. Arnaldo 455, Cerqueira César, 01246-903, S?o Paulo, SP, Brazil
3. Mathematic and Statistic Institute, S?o Paulo University, Cidade Universitária Rua do Mat?o 1010, Cidade Universitária, 05508-000, S?o Paulo, SP, Brazil
4. Dental School Pathology Department, S?o Paulo University, Av. Lineu Prestes 2227, Cidade Universitária, 05508-000, S?o Paulo, SP, Brazil
Abstract:The possibility of obtaining transplantable oral epithelia opens new perspectives for oral treatments. Most of them are surgical, resulting in mucosal failures. As reconstructive material this in vitro epithelia would be also useful for other parts of the human body. Many researchers still use controversial methods; therefore it was evaluated and compared the efficiency of the enzymatic and direct explant methods to obtain oral keratinocytes. To this project oral epithelia fragments were used. This work compared: time needed for cell obtainment, best cell amount, life-span and epithelia forming cell capacity. The results showed the possibility to obtain keratinocytes from a small oral fragment and we could verify the advantages and peculiar restrictions. We concluded that under our conditions the enzymatic method showed the best results: in the cells obtaining time needed, cell amount and life-span. Both methods showed the same capacity to form in vitro epithelia.
Keywords:Primary cell culture  Keratinocytes  Oral keratinocytes  Cell culture techniques  Primary culture techniques
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