RESUMEN
Objective To explore the cultural method of lingual keratinocytes and make preparations for further investigation in using lingual keratinocytes as a new choice of seed cells for the reconstruction of tissue engineered corneal epithelium.Methods Keratinocytes were enzymatically dissociated from lingual tissue specimens using 0.25%disepase and 0.05%trypsin.The biological characteristics of the cells were observed through phase microscope.Meanwhile.1ingual kemtinoeytes were immtmohistochemically labeled with broad spectrum keratin antibody (AE1/AF3).Results The cells revealed the form of "road metal" and could be passed for 3 or 4 generations.Immunohistochemistry indicated that lingual keratinocytes were positive for AE1/AE3 staining.Condusion Lingual kemtinocytes of rabbits can be cultured with KSFM in vitm and magnitude quantity can be attained,laying a favorable foundation for lingual keratinocytes as a new choice of seed cells for corneal epithelium reconstruction with tissue engineering.
RESUMEN
Objective To investigate the effects of imiquimod on cytokine secretions of normal human epidermal Langerhans cells (LCs) and HaCaT cells, and to better understand the mechanism of immune regulation by imiquimod. Methods LCs were sorted from prepared epidenmal cells by density gradient centrifugation and magnetic-activated cell sorting (MACS). LCs and HaCaT cells were cultured in media with or without 5 ?g/mL of imiquimod for 4 hours, then cell-free culture supernatants were harvested, cytokines were detected by ELISA kits. Results TNF-?, IL-1? and IL-6 secreted from imiquimod treated LCs were all higher than those from control LCs (all P