A perfusable 3D cell-matrix tissue culture chamber for in situ evaluation of nanoparticle vehicle penetration and transport
A perfusable 3D cell-matrix tissue culture chamber for in situ evaluation of nanoparticle vehicle penetration and transport
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DOI：
10.1002/bit.21698
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发表时间：
2008-04-15
影响因子：
3.8
通讯作者：
Pun, Suzie Hwang
中科院分区：
文献类型：
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作者：
Ng, Chee Ping;Pun, Suzie Hwang
基因或药物治疗中的一个关键因素是开发能够有效地从远端给药到达靶细胞的载体。在许多基因/药物传递研究中，在培养中获得的结果在体内无法转化为类似的结果。在这项工作中，我们开发了一种可灌流的小室，用于研究纳米粒子在细胞凝胶软组织培养物中的渗透和传输。隔室由聚二甲基硅氧烷(PDMS)顶层制成，具有腔体特征，使用微机械光刻技术创建，粘接在底部玻璃盖板上。将嵌入水凝胶中的细胞溶液加载到PDMS柱之间的腔室中，PDMS柱在凝胶-介质界面处作为细胞基质的锚。该试验室具有以下独特特点：(I)快速制造和组装简单，(Ii)在垂直于流动或作用方向的平面上直接进行原位细胞成像，(Iii)易于配置和控制的环境，有利于静态或间质流动条件下的细胞培养，以及(Iv)从试验室中轻松回收活细胞用于实验后分析。为了评估小室，我们将三种不同大小的荧光标记纳米颗粒输送到细胞-在流动和静态条件下嵌入3D小室中的Matrigels。在间质流动下，纳米颗粒的渗透性得到了增强，而活细胞成像和回收细胞的流式细胞仪显示，纳米颗粒的大小限制了有效的传递。尽管是为分娩研究而设计的，但这个试验室是多功能的，可以很容易地进行改装。因此，它可能在生物、组织工程和治疗研究中有广泛的应用。
A key factor in gene or drug therapy is the development of carriers that can efficiently reach targeted cells from a distal administration. In many gene/drug delivery studies, results obtained in 21) cultures fail to translate to similar results in vivo. In this work, we developed a perfusable 31) chamber for studying nanoparticle penetration and transport in cell-gel soft tissue cultures. The compartmented chamber is made of a polydimethylsiloxane (PDMS) top layer with the chamber features, created using micromachined lithography, bonded to a bottom glass coverslip. A solution of cells embedded in a hydrogel is loaded in the chamber between PDMS posts that serve as anchors to the cell-matrix at the gel-media interface. The chamber offers the following unique features: (i) rapid fabrication and simplicity in assembly, (ii) direct in situ cell imaging in a plane normal to the direction of flow or action, (iii) an easily configurable and controllable environment conducive cell culture under static or interstitial flow conditions, and (iv) facile recovery of live cells from chambers for post-experimental analysis. To assess the chamber, we delivered fluorescently labeled nanoparticles of three distinct sizes to cells-embedded Matrigels in the 3D chamber under flow and static conditions. Penetration of nanoparticles were enhanced under interstitial flow while live cell imaging and flow cytometry of recovered cells revealed particle size restrictions to efficient delivery. Although designed for delivery studies, the chamber is versatile and can be easily modified. Thus it may have broad applications for biological, tissue engineering, and therapeutic studies.