@misc{oai:niigata-u.repo.nii.ac.jp:00005378, author = {Monir, Mar Zabin Binta}, month = {Sep}, note = {Fibroblasts in monoculture grown in serum-containing medium are highly-proliferating. However, data suggested that use of static fibroblasts provides a better model to study biological phenomena than proliferating fibroblasts even when constructing a three-dimensional matrix. Since previous studies have stated keratinocyte-conditioned medium reduced fibroblast proliferation, this study aimed to examine if we can harness the discarded oral keratinocyte-conditioned medium (OK-CM) for oral fibroblasts (OF) culture in a monolayer and to investigate their phenotypic changes by OK-CM from an actively-cycling state to a more static state as well as their behavior. Primary human OK and OF were grown in a chemically-defined, completed EpiLife and DMEM containing 10% calf serum (DMEM-CS), respectively. OK-CM conditioned for 24 hours was obtained from near-confluent OK culture. OF were plated into microplate-wells, and 24 hours later, the medium was replaced with DMEM-CS, serum-free DMEM (SF-DMEM) and OK-CM. OF were cultured up to 96 hours. Proliferation rate and cell cycle profile were analyzed using a MTT assay and a fluorescence-activated cell sorter. To assess the phenotypic changes of OFs cultured in each medium, the activity of senescent-associated β-GAL and the secreted protein levels including Keratinocyte growth factor (KGF), human type I collagen and MMP-1 were determined. The proliferating rate and the proportion of cells in S and G2/M significantly decreased when cells were cultured in OK-CM, indicating the inhibition of cell cycle progression at the G1 phase. In contrast to OFs in SF-DMEM, only a few OFs cultured in OK-CM showed the β-gal activity, suggesting that they had proliferating potential. ELISA assay showed OFs cultured in OK-CM produce KGF and MMP-1 as do OFs grown in DMEM-CS. However, their ability to produce type I collagen significantly decreased compared with OFs in DMEM-CS. Significant OK-CM-mediated OF phenotypic changes suggest the feasibility of those quiescent OFs cultured in OK-CM for reconstruction of a three-dimensional tissue-engineered oral mucosa substitute., 学位の種類: 博士(歯学). 報告番号: 甲第3816号. 学位記番号: 新大院博(歯)甲第282号. 学位授与年月日: 平成25年9月20日, 新大院博(歯)甲第282号}, title = {Characterization of an oral fibroblast phenotype cultured in oral keratinocyte-conditioned medium}, year = {2013} }