Construction and harvest of multilayered keratinocyte sheets using magnetite nanoparticles and magnetic force
Construction and harvest of multilayered keratinocyte sheets using magnetite nanoparticles and magnetic force
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DOI：
10.1089/1076327041348446
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发表时间：
2004-05-01
影响因子：
--
通讯作者：
Kobayashi, T
中科院分区：
文献类型：
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作者：
Ito, A;Hayashida, M;Kobayashi, T
建立三维结构的新技术是组织工程所需要的。在本研究中，使用磁力来构建多层角质形成细胞片层，并在没有酶处理的情况下收获片层。我们最初的磁铁矿阳离子脂质体具有正表面电荷以改善吸附，其被人角质形成细胞以33 pg磁铁矿/细胞的浓度吸收。将磁性标记的角质形成细胞（2 × 10（6）个细胞，其对应于24孔板的培养区域的5倍汇合浓度，以产生5层角质形成细胞片层）接种到24孔超低附着板中，其表面由亲水性和中性电荷的共价结合的水凝胶层组成。将磁体（4000 G）置于孔下，培养24小时后，角质形成细胞在低钙培养基（钙浓度，0.15 mM）中形成五层结构。随后，当在高钙培养基（钙浓度，1.0mM）中培养五层角质形成细胞时，角质形成细胞进一步分层，导致形成10层表皮片。当移除磁体时，将片材从板的底部分离，并且可以用磁体收获片材。这些结果表明，这种新的方法，使用磁铁矿纳米粒子和磁力，我们称之为“基于磁力的组织工程”（Mag-TE），是一种很有前途的组织工程方法。
Novel technologies to establish three-dimensional constructs are desired for tissue engineering. In the present study, magnetic force was used to construct multilayered keratinocyte sheets and harvest the sheets without enzymatic treatment. Our original magnetite cationic liposomes, which have a positive surface charge in order to improve adsorption, were taken up by human keratinocytes at a concentration of 33 pg of magnetite per cell. The magnetically labeled keratinocytes (2 x 10(6) cells, which corresponds to 5 times the confluent concentration against the culture area of 24-well plates, in order to produce 5-layered keratinocyte sheets) were seeded into a 24-well ultralow-attachment plate, the surface of which was composed of a covalently bound hydrogel layer that is hydrophilic and neutrally charged. A magnet ( 4000 G) was placed under the well, and the keratinocytes formed a five-layered construct in low-calcium medium ( calcium concentration, 0.15 mM) after 24 h of culture. Subsequently, when the five-layered keratinocytes were cultured in high-calcium medium ( calcium concentration, 1.0 mM), keratinocytes further stratified, resulting in the formation of 10-layered epidermal sheets. When the magnet was removed, the sheets were detached from the bottom of the plates, and the sheets could be harvested with a magnet. These results suggest that this novel methodology using magnetite nanoparticles and magnetic force, which we have termed "magnetic force-based tissue engineering" (Mag-TE), is a promising approach for tissue engineering.