LIF-immobilized nonwoven polyester fabrics for cultivation of murine embryonic stem cells
LIF-immobilized nonwoven polyester fabrics for cultivation of murine embryonic stem cells
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DOI：
10.1002/jbm.a.31107
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发表时间：
2007-06-15
影响因子：
4.9
通讯作者：
Tumer, Askin
中科院分区：
文献类型：
--
作者：
Cetinkaya, Gaye;Turkoglu, Hilal;Tumer, Askin
胚胎干细胞（ES细胞）由于其多能性和增殖能力而在组织工程中引起了极大的兴趣。本研究的目的是建立一个合成的微环境，以支持在体外繁殖的小鼠ES细胞在未分化状态。这就是为什么我们使用由聚对苯二甲酸乙二醇酯（PET）纤维制成的三维基质非织造聚酯织物（NWPF）。将NWPF片部分水解，然后在水溶性碳二亚胺存在下将羧基与白血病抑制因子（LIF）偶联。固定化过程的有效性进行了检查与ATR-FTIR光谱，荧光，和细胞培养研究。ES细胞集落形态，碱性磷酸酶（AP）的活性，阶段特异性胚胎抗原-1（SSEA-1）的免疫反应性，并在水解和LIF固定的表面上培养72-96小时后的SEM分析进行了评估，以确定ES细胞的多能性状态。结果表明，LIF是活跃的固定化形式，未分化的菌落不仅有显着的AP和SSEA-1的免疫反应性，但也有较高的未分化的集落比例LIF固定化表面比水解表面。固定化的LIF蛋白可能是一个很好的模型，以提供一个无饲养系统，但支架的物理性质是更方便的分化研究。(c)2007 Wiley Periodicals，Inc. J Biomed Mater Res 81 A：911-919，2007.
Embryonic stem (ES) cells have a great interest for tissue engineering because of their pluripotent nature and proliferative capacity. The objective of this study is to constitute a synthetic microenvironment to support the in vitro propagation of murine ES cells in an undifferentiated state. That is why we used a three-dimensional matrix, nonwoven polyester fabric (NWPF), which was formed from poly(ethylene terephthalate) (PET) fibers. NWPF discs were partially hydrolyzed, and then the carboxyl groups were coupled with leukemia inhibitory factor (LIF) in the presence of water-soluble carbodiimide. The effectiveness of immobilization process was checked with ATR-FTIR spectroscopy, fluorimetry, and cell culture studies. ES cell colony morphology, alkaline phosphatase (AP) activity, stage-specific embryonic antigen-1 (SSEA-1) immunoreactivity, and SEM analysis following a 72-96-h culture period upon hydrolyzed and LIF-immobilized surfaces were assessed to determine the pluripotent status of ES cells. Results revealed that LIF was active in immobilized form; undifferentiated colonies had not only a significant AP and SSEA-1 immunoreactivity, but also a higher undifferentiated colony ratio on LIF-immobilized surfaces than that of hydrolyzed surfaces. The immobilized LIF protein might be a good model to provide a feeder-free system, but the physical properties of the scaffold is more convenient for differentiation studies. (c) 2007 Wiley Periodicals, Inc. J Biomed Mater Res 81A: 911-919, 2007.