Sakai Shinji, Ogushi Yuko, Kawakami Koei
Department of Chemical Engineering, Faculty of Engineering, Kyushu University, 744 Motooka, Nishi-ku, Fukuoka 819-0395, Japan.
Acta Biomater. 2009 Feb;5(2):554-9. doi: 10.1016/j.actbio.2008.10.010. Epub 2008 Oct 30.
An aqueous solution of carboxylmethylcellulose with phenolic hydroxyl groups (CMC-Ph) is gellable within 1 min via a peroxidase-catalyzed oxidative reaction under mild conditions suitable for mammalian cells. In this research, we evaluated cellular adhesion and proliferation on the resultant hydrogel, and the feasibility of the hydrogel as a substrate for cell sheet technology. Within 4 h of seeding, 76.9% of L929 fibroblast cells adhered to the gel and showed similar morphology of spreading to that on cell culture dish. Subsequently, the adherent cells proliferated on the gel and formed a confluent monolayer after 168 h of culture. From the confluent monolayer we could harvest a cell sheet after about 5 min of digestion of the gel using cellulase dissolved in medium at 5 U ml(-1). The cells in the cell sheet showed well-preserved morphology similar to that shown before they were detached from the gel. In addition, the harvested cell sheet readhered and proliferated after being transferred to another culture dish. These results demonstrate that CMC-Ph gel is a good candidate material for obtaining cell sheets.
含酚羟基的羧甲基纤维素（CMC-Ph）水溶液在适合哺乳动物细胞的温和条件下，可通过过氧化物酶催化的氧化反应在1分钟内形成凝胶。在本研究中，我们评估了细胞在所得水凝胶上的黏附与增殖情况，以及该水凝胶作为细胞片技术底物的可行性。接种后4小时内，76.9%的L929成纤维细胞黏附于凝胶上，且呈现出与在细胞培养皿上类似的铺展形态。随后，黏附的细胞在凝胶上增殖，并在培养168小时后形成汇合的单层细胞。从汇合的单层细胞中，使用溶解于培养基中的5 U ml(-1)纤维素酶消化凝胶约5分钟后，我们能够收获细胞片。细胞片中的细胞呈现出保存良好的形态，与从凝胶上脱离前的形态相似。此外，收获的细胞片转移至另一培养皿后能够重新黏附并增殖。这些结果表明，CMC-Ph凝胶是获取细胞片的良好候选材料。
