A novel culture system shows that stem cells can be grown in 3D and under physiologic pulsatile conditions for tissue engineering of vascular grafts
A novel culture system shows that stem cells can be grown in 3D and under physiologic pulsatile conditions for tissue engineering of vascular grafts
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DOI：
10.1016/j.jss.2006.02.017
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发表时间：
2006-05-15
影响因子：
2.2
通讯作者：
Zarins, C
中科院分区：
文献类型：
--
作者：
Abilez, O;Benharash, P;Zarins, C
背景目前可用的血管移植物受到可变的通畅率、材料可用性和免疫排斥的限制。从自体干细胞中产生组织工程血管移植物（TEVG）可能会克服这些限制。作为创建完全自体TEVG的第一步，我们的目标是开发一种用于在三维（3D）配置中以及在生理脉动流和压力条件下培养未分化小鼠胚胎干细胞（mESC）的新系统。创建生物反应器以向专门修改的四孔Labtek Chamber-Slide培养系统提供脉动条件。将未分化的mESC悬浮在3D Matrigel基质中或仅悬浮在培养系统内的细胞培养基中。将脉动条件应用于悬浮细胞并通过视频显微镜观察。未分化的mESC被成功地包埋在3D Matrigel基质中，并且可以承受生理脉动条件。视频显微镜显示，三维基质中的mESC被限制在培养系统的威尔斯孔中，与施加的流动一致地移动，并且不被下游冲洗;这与单独悬浮在培养基中的mESC形成对比。未分化的mESC可以在3D和脉动条件下生长。我们将使用这些结果来研究长期脉动条件对mESC分化为内皮细胞、平滑肌细胞和成纤维细胞的影响，长期目标是创造完全自体的TEVG。(c)2006年爱思唯尔公司All rights reserved.
Background. Currently available vascular grafts have been limited by variable patency rates, material availability, and immunological rejection. The creation of a tissue-engineered vascular graft (TEVG) from autologous stem cells would potentially overcome these limitations. As a first step in creating a completely autologous TEVG, our objective was to develop a novel system for culturing undifferentiated mouse embryonic stem cells (mESC) in a three-dimensional (3D) configuration and under physiological pulsatile flow and pressure conditions.Materials and methods. A bioreactor was created to provide pulsatile conditions to a specially modified four-well Labtek Chamber-Slide culture system. Undifferentiated mESC were either suspended in a 3D Matrigel matrix or suspended only in cell-culture media within the culture system. Pulsatile conditions were applied to the suspended cells and visualized by video microscopy.Results. Undifferentiated mESC were successfully embedded in a 3D Matrigel matrix and could withstand physiological pulsatile conditions. Video microscopy demonstrated that the mESC in the 3D matrix were constrained to the wells of the culture system, moved in unison with the applied flows, and were not washed downstream; this was in contrast to the mESC suspended in media alone.Conclusions. Undifferentiated mESC can be grown in 3D and under pulsatile conditions. We will use these results to study the effects of long-term pulsatile conditions on the differentiation of mESC into endothelial cells, smooth muscle cells, and fibroblast cells with the long-term goal of creating a completely autologous TEVG. (c) 2006 Elsevier Inc. All rights reserved.