@article{oai:niigata-u.repo.nii.ac.jp:02000494,
 author = {モニル, マザビンビンタ and Monir, Mah Zabin Binta},
 issue = {2},
 journal = {新潟歯学会雑誌},
 month = {Dec},
 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. Since previous studies have stated keratinocyte-conditioned medium reduced fibroblast proliferation, this study aimed to examine if the oral keratinocyte (OK) -conditioned medium (CM) can decrease oral fibroblasts (OF) proliferation and to characterize their phenotype. Primary human OK and OF were grown in a completed EpiLife^<【○!R】>(0.06mM Ca^<++>) and Dulbecco's modified Eagle medium (DMEM) containing 10% calf serum (DMEM-CS), respectively. OK-CM was conditioned for 24 hours in a near-confluent OK culture. OFs plated in a micro-plate well were cultured with DMEM-CS, serum-free DMEM (SF-DMEM) and OK-CM for up to 96 hours. Proliferation rate and cell cycle profile were analyzed using a MTT assay and a fluorescence-activated cell sorter. The "phenotypic changes" of OFs were determined by the activity of senescent-associated β-galactosidase (β-gal) and the secreted protein levels including keratinocyte growth factor (KGF), human type I collagen and matrix metalloproteinase-1 (MMP-1) measured by enzyme-linked immunosorbent assay (ELISA). The proliferating rate and the proportion of cells in S phase were significantly lower when cells were cultured in OK-CM. The β-gal activity suggested OFs in OK-CM still had proliferating potential. ELISA assay showed OFs cultured in OK-CM produce KGF and MMP-1 as did OFs grown in DMEM-CS while their ability to produce type I collagen was significantly lower than OFs in DMEM-CS. This study suggested the OK-CM generated a quiescent OF population possessing the characteristic of extracellular matrix degradation rather than synthesis., 生体反応の研究に用いる線維芽細胞は増殖している細胞ではなく休止期のものが好ましい。上皮細胞の培養上清に線維芽細胞の増殖抑制効果があることが報告されている。本研究の目的は口腔粘膜上皮細胞の培養上清が培養口腔粘膜線維芽細胞の増殖性を抑え，細胞形質を変化させるかを検証することである。ヒト初代口腔粘膜上皮細胞(OK)と線維芽細胞(OF)はそれぞれEpiLifeと10%ウシ胎児血清含有ダルベッコ改変イーグル培地(DMEM)(DMEM-CS)中で培養した。コンフルエントに近い口腔粘膜上皮細胞を24時間培養したものを上清とした。口腔粘膜線維芽細胞はDMEM-CS，DMEM培地単独(SF-DMEM)，口腔粘膜上皮細胞培養上清(OK-CM)の3種類の培地で，最長96時間培養した。細胞増殖能と細胞周期はそれぞれMTT assayとセルソーターで解析した。細胞形質変化の分析はさらに，βガラクトシダーゼ活性発現とエライザ法(ELISA)による各種タンパク質(ケラチノサイト増殖因子(KGF)，タイプIコラーゲン，マトリックスメタロプロテアーゼ-1(MMP-1))分泌量で検討した。細胞増殖能とS期の細胞の割合は有意にOK-CMで培養した細胞で低下しており，G0/G1期停止による細胞増殖抑制の可能性が示唆された。またOK-CM培養では，βガラクトシダーゼ活性を示した細胞は少数で，口腔粘膜線維芽細胞は休止期にいるが，増殖能が失われていないことを示した。KGFとMMP-1の産生量は，DMEM-CSで培養した細胞と同等であったのに対し，I型コラーゲンの産生量が有意に低かった。本研究から，OK-CMは培養口腔粘膜線維芽細胞の増殖能を抑え，かつ細胞外基質分解性の細胞形質となることが示唆された。},
 pages = {105--112},
 title = {口腔粘膜上皮細胞培養上清で培養した口腔粘膜線維芽細胞の形質に関する検討},
 volume = {43},
 year = {2013}
}