Control of cellular adhesiveness in an alginate-based hydrogel by varying peroxidase and H(2)O(2) concentrations during gelation.
Control of cellular adhesiveness in an alginate-based hydrogel by varying peroxidase and H(2)O(2) concentrations during gelation.
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DOI：
10.1016/j.actbio.2009.10.004
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发表时间：
2010-04
影响因子：
9.7
通讯作者：
S. Sakai;K. Hirose;Kousuke Moriyama;K. Kawakami
中科院分区：
文献类型：
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作者：
S. Sakai;K. Hirose;Kousuke Moriyama;K. Kawakami
具有酚羟基（Alg-Ph）基团的海藻酸盐的水溶液可通过辣根过氧化物酶（HRP）催化的Ph基团之间的氧化交联反应而凝胶化，消耗H2 O2作为电子受体。本研究评估了H2 O2和HRP浓度对所得酶交联Alg-Ph凝胶上细胞增殖和增殖的影响。接种4小时后，81.1%的L929成纤维细胞粘附到用1Uml−1HRP和1 mM H2 O2制备的Alg-Ph水凝胶上。当H_2O_2浓度增加到15 mM时，细胞粘附率下降到28.4%。在此H2 O2浓度下，通过将HRP浓度增加至10 Uml −1，细胞粘附增加至82.6%。在168 h的培养过程中，粘附在具有较高细胞粘附性的Alg-Ph水凝胶上的细胞建立了汇合的单层。然后可以在浸入含有1.0mgml-1藻酸盐裂解酶的培养基中5分钟内收获细胞片层。收获的细胞片重新粘附，并且片中包含的细胞在转移到另一个细胞培养皿后增殖。
An aqueous solution of alginate possessing phenolic hydroxyl (Alg-Ph) groups is gellable via a horseradish peroxidase (HRP)-catalyzed oxidative crosslinking reaction between Ph groups, consuming H2O2as an electron acceptor. This study evaluates the effect of H2O2and HRP concentrations on cellular adhesiveness and proliferation on the resultant enzymatically crosslinked Alg-Ph gels. After 4h of seeding, 81.1% of L929 fibroblast cells adhere to an Alg-Ph hydrogel prepared with 1Uml−1HRP and 1mM H2O2. Increasing the concentration of H2O2to 15mM decreases the percentage of adhering cells to 28.4%. The cellular adhesion at this H2O2concentration is increased to 82.6% by increasing the HRP concentration to 10Uml−1. The cells adhering to the Alg-Ph hydrogels with higher cellular adhesiveness establish a confluent monolayer during 168h of culture. A cell sheet can then be harvested within 5min of immersion in a medium containing alginate lyase at 1.0mgml−1. The harvested cell sheet re-adhere, and the cells contained in the sheet proliferate after being transferred to another cell culture dish.