Horseradish peroxidase/catalase-mediated cell-laden alginate-based hydrogel tube production in two-phase coaxial flow of aqueous solutions for filament-like tissues fabrication
Horseradish peroxidase/catalase-mediated cell-laden alginate-based hydrogel tube production in two-phase coaxial flow of aqueous solutions for filament-like tissues fabrication
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DOI：
10.1088/1758-5082/5/1/015012
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发表时间：
2013-01
期刊：
影响因子：
9
通讯作者：
S. Sakai;Yang Liu;Emma J. Mah;M. Taya
中科院分区：
文献类型：
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作者：
S. Sakai;Yang Liu;Emma J. Mah;M. Taya
我们报道了一种制备载细胞水凝胶管的方法。该方法使用同轴双孔喷丝器，比用于制备类似结构的三孔喷丝器更简单。目的是创建一个模板，用于制备由两个异质活细胞层组成的丝状结构。将含有含有酚羟基部分的藻酸盐衍生物（Alg-Ph）、过氧化氢酶和辣根过氧化物酶（HRP）的水溶液挤出H2O2水溶液中。该操作使Alg-Ph溶液通过酶催化反应可凝胶化，将Ph部分交联在一起。通过改变Alg-Ph和H2O2溶液的流速以及过氧化氢酶和H2O2的浓度，水凝胶管的直径和膜厚度分别在250 ~ 550µm和70 ~ 140µm之间可控。在直径为300µm、膜厚为80µm的水凝胶管中，HeLa细胞的存活率为95.4%。随后，封闭的海拉细胞生长并填满空心核心。当成纤维细胞10T1/2细胞达到合流状态，用海藻酸解酶降解水凝胶基质后，得到了HeLa细胞表面覆盖一层成纤维细胞10T1/2的丝状结构。
We report a method for preparing cell-laden hydrogel tubes. This method uses a coaxial double-orifice spinneret, simpler than triple-orifice spinnerets which have been used for preparing similar constructs. The intended application was to create a template for preparing filament-like structures composed of two heterogeneous living cell layers. An aqueous solution containing an alginate derivative possessing phenolic hydroxyl moieties (Alg-Ph), catalase and horseradish peroxidase (HRP) was extruded into an ambient flow of H2O2 aqueous solution. This operation enabled the Alg-Ph solution to be gellable through a HRP-catalyzed reaction, cross-linking the Ph moieties together. By altering flow rates of the Alg-Ph and H2O2 solutions along with the concentrations of catalase and H2O2, the diameter and membrane thickness of the hydrogel tubes were controllable between 250–550 µm and 70–140 µm, respectively. The viability of the HeLa cells enclosed in the hydrogel tubes with a diameter of 300 µm and a membrane thickness of 80 µm was 95.4%. Subsequently, the enclosed HeLa cells grew and filled the hollow core. A filament-like structure of HeLa cells covered with a layer of fibroblast 10T1/2 cells was obtained when confluency of fibroblast 10T1/2 cells was reached and the hydrogel matrix was degraded with alginate lyase.