(1) 各妊娠時期での羊膜マトリクス細胞蛋白の発現変化 : SPARC mRNA発現は, 妊娠後期に40%減少し, TSP1は3倍に増加した。(2) 羊膜プロスタグランジン合成酵素の発現変化 : COX2 mRNAのみが妊娠後期に2倍に増加した。(3) 流動細胞計測法を用いた羊膜上皮細胞分離法の確立 : 当初の上皮細胞増殖が不安定で, 実験計画の変更を余儀なくされたため, 新たな分離法を確立した。(4) 羊膜上皮細胞における網羅的遺伝子発現解析 : 初代培養で増殖因子のmidkineやFGF2発現が低く, 免疫抑制因子HLA-G発現は高かった。継代後midkine, FGF2の発現は急速に増加し, HLA-G 発現は減少した。
(1) Expression changes of matrix cellular proteins of the amniotic membrane in mid and late gestation : the expression of SPARC mRNA was reduced by 40% at the late gestation. Expression of TSP1 was increased three-fold in late gestation. (2) Expression changes of prostaglandin synthesis enzyme : only COX2 mRNA expression was increased twice at the late gestation. (3) Establishment of amniotic epithelial cell separation method using flow cytometry : because there was instability in the cell growth of the original amniotic epithelial cells, it was forced to change the experimental plan. A new separation method was established. (4) Comprehensive gene expression analysis in epithelial cells obtained by flow cytometry : in primary epithelial cells, the expression of midkine and FGF2 involved in the proliferation was low, the expression of HLA-G related to immunosuppression was higher. After passage, the expression of midkine and FGF2 was increased rapidly, HLA-G expression was decreased.
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