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Cell Journal، جلد ۱۶، شماره Suppl ۱، صفحات ۱۶۸-۱۶۸
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عنوان انگلیسی Ps-137: Reducing The Risks in Producing Tissue Engineered Buccal Mucosa for Clinical Use
چکیده انگلیسی مقاله Objective: Tissue engineered (TE) buccal mucosa has been used with good clinical outcomes in reconstructive surgery for the urethra. This involved the use of human acellular donor dermis, murine 3T3 fibroblasts as a feeder layer to expand oral keratinocytes and bovine foetal calf serum (FCS) to provide mitogens for these cells. Our aim was to avoid the use of donor human material, animal derived cells and sera in the production of TE oral mucosa, making it safer for clinical use. Our objectives accordingly were to replace human donor dermis with a biodegradable electrospun scaffold to be used as a synthetic dermal alternative and to replace mouse fibroblasts with screened human fibroblast as a feeder layer to culture oral keratinocytes and to avoid the use of FCS. Materials and Methods: 10% PolyL-lactide (PLLA) in dichloromethane was electrospun to produce a scaffold. The human fetal lung fibroblast cell line MRC-5 was used to replace murine 3T3J2 fibroblasts or human oral fibroblasts were compared. In all cases oral keratinocytes were seeded into the scaffolds either in the presence or absence of FCS. In separate experiments media was supplemented with basic fibroblast growth factor (bFGF) or ascorbate-2-phosphate to increase collagen production. The results were assessed using AlamarBlue for cell viability and Sirius red to assess collagen production on days 7 and 14.Results: Cells grew well on a 10% PLLA scaffold. We were able to expand oral keratinocytes in serum free conditions using either oral fibroblasts or MRC-5 fibroblasts as a feeder layer. The presence of bFGF or ascorbic acid-2-phosphate also increased collagen production. Conclusion: We have achieved several steps to produce TE buccal mucosa which does not require donor human tissue, murine feeder cells or FCS and thus presents less risk of viral disease transmission for the patient.
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نشانی اینترنتی http://celljournal.org/journal/article/abstract/462
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