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ジャイアントリポソーム融合による生細胞へのマイクロメートルスケール人工物の導入

通过巨型脂质体融合将微米级人工制品引入活细胞

基本信息

批准号：
15J02547
负责人：
齋藤明
金额：
$ 1.47万
依托单位：
Tohoku University
依托单位国家：
日本
项目类别：
Grant-in-Aid for JSPS Fellows
财政年份：
2015
资助国家：
日本
起止时间：
2015-04-24 至 2017-03-31
项目状态：
已结题

项目摘要

昨年度は，望みの巨大物質を内包させたGUV(巨大脂質二重膜リポソーム)を標的細胞と電気融合させることによってGUVから細胞へとマイクロメートルサイズの物質を導入する手法を確立した．本年度はこの結果を基に，細胞とGUVの数比率やGUV膜の組成を変化させることで，細胞への物質の導入効率がどのように影響するか調査し，導入効率の向上を図り，最適条件について検討した．また異なる細胞種を用いることで，本法による導入効率の細胞種依存性についても調査し，同じく細胞への導入可能物質についても調査した．具体的には，細胞とマイクロサイズのビーズを内包したGUVの比率を調整して導入効率を評価した．その結果，細胞に対するGUVの割合が多いほど，物質の導入率が向上することが明らかになっ．また，GUV脂質膜の組成と細胞への物質導入率の影響について調査するため，脂質膜表面電荷の異なるGUV(正，中性，負)を用いて電気融合を行い，導入率を評価した．その結果，正電荷及び負電荷脂質膜を使用して融合した場合に導入率が格段に向上することが明らかになった．加えて，細胞自体のサイズと人工物導入率の関連について調査した結果，細胞側の直径が大きいほど導入率が向上し，導入可能なサイズの上限も増加するという結果が得られた．さらに，導入可能な物質を拡大するべく調査を行った．その結果，活性を保ったまま抽出し，生細胞へ導入するのが困難とされている細胞内小器官であるミトコンドリアを，本法を用いて活性を保ちつつ細胞内に導入できることが明らかになった．以上，本年度の研究によって，本法における細胞サイズと導入可能物質との関連について従来不明であった新たな知見が得られたと共に，導入率をより向上させるパラメータが見出されたといえる．

Last year, it was hoped that the cell phone of the giant fat double membrane giant GUV (giant lipid double membrane) was fused with the cell phone of the giant lipid double membrane (GUV). The cell of the giant fat double membrane (GUV) was fused with the cell of the GUV (giant lipid double membrane). This year, the results of the experiment are based on the ratio of cell GUV to the number of cells. The input rate of the cell is different from that of the control. The input rate is higher than that of the control group, and the most suitable condition is that the input rate of the cell is higher than that of the control. This method applies this method to the cell-dependent property of the cell, which is different from that of the other cell. The percentage of GUV in the cell is different from that in the control group. The results show that the input rate of the GUV is higher than that of the control group, and the input rate of the GUV lipid membrane is higher than that of the control group. The GUV lipid membrane is composed of a negative temperature, and a positive, neutral surface charge temperature. In the end, the positive charge and the loading rate of the lipid membrane were significantly higher than those of the control group. In addition, the input rate of artificial materials was higher than that of the control group, and the cell diameter was higher than that of the others. The results show that there are significant differences between the two groups. The results show that there are significant differences between the two groups. The results show that there are significant differences between the two groups. The results show that there are significant differences between the two groups. The results show that there is a significant increase in the number of small organs in the cell. the results show that there is a significant increase in the number of cells. This method is based on the activity of the cell. This method is based on the fact that this year's research has been carried out in this year, and this method has been used in this year's research. This method is based on the study of the possibility that new information can be obtained in this way, and the rate of admission will increase.