Stephens J S, Cooper J A, Phelan F R, Dunkers J P
Polymers Division, National Institute of Standards and Technology, 100 Bureau Dr, Gaithersburg, Maryland 20899, USA.
Biotechnol Bioeng. 2007 Jul 1;97(4):952-61. doi: 10.1002/bit.21252.
The capability to image real time cell/material interactions in a three-dimensional (3D) culture environment will aid in the advancement of tissue engineering. This paper describes a perfusion flow bioreactor designed to hold tissue engineering scaffolds and allow for in situ imaging using an upright microscope. The bioreactor can hold a scaffold of desirable thickness for implantation (>2 mm). Coupling 3D culture and perfusion flow leads to the creation of a more biomimetic environment. We examined the ability of the bioreactor to maintain cell viability outside of an incubator environment (temperature and pH stability), investigated the flow features of the system (flow induced shear stress), and determined the image quality in order to perform time-lapsed imaging of two-dimensional (2D) and 3D cell culture. In situ imaging was performed on 2D and 3D, culture samples and cell viability was measured under perfusion flow (2.5 mL/min, 0.016 Pa). The visualization of cell response to their environment, in real time, will help to further elucidate the influences of biomaterial surface features, scaffold architectures, and the influence of flow induced shear on cell response and growth of new tissue.
在三维（3D）培养环境中对实时细胞/材料相互作用进行成像的能力将有助于组织工程的发展。本文描述了一种灌注流生物反应器，其设计用于容纳组织工程支架，并允许使用直立显微镜进行原位成像。该生物反应器可以容纳适合植入的厚度（>2毫米）的支架。将3D培养与灌注流相结合可营造出更具仿生学的环境。我们研究了生物反应器在培养箱外环境中维持细胞活力的能力（温度和pH稳定性），研究了系统的流动特性（流动诱导剪切应力），并确定了图像质量，以便对二维（2D）和3D细胞培养进行延时成像。对2D和3D培养样本进行原位成像，并在灌注流（2.5毫升/分钟，0.016帕）下测量细胞活力。实时观察细胞对其环境的反应将有助于进一步阐明生物材料表面特征、支架结构以及流动诱导剪切对细胞反应和新组织生长的影响。
