The construction of 3D-engineered tissues composed of cells and extracellular matrices by hydrogel template approach
The construction of 3D-engineered tissues composed of cells and extracellular matrices by hydrogel template approach
复制标题
DOI：
10.1016/j.biomaterials.2007.02.015
复制
发表时间：
2007-06-01
期刊：
影响因子：
14
通讯作者：
Akashi, Mitsuru
中科院分区：
文献类型：
--
作者：
Matsusaki, Michiya;Yoshida, Hiroaki;Akashi, Mitsuru
通过水凝胶模板法构建了仅由细胞和细胞外基质（ECM）组成的三维（3D）工程组织。制备了二硫键交联的聚（γ-谷氨酸）水凝胶作为模板水凝胶。这些模板水凝胶在生理条件下使用还原剂如半胱氨酸、谷胱甘肽和二硫苏糖醇通过将二硫键交联裂解成硫醇基团而容易地分解。分解的聚合物可溶于细胞培养基。用紫外可见光谱和红外光谱研究了二硫键的断裂。我们成功地制备了由小鼠L929成纤维细胞和ECM组成的3D工程组织（厚度/直径，2 mm/1cm），通过在模板水凝胶上/中3D细胞培养10天后仅用半胱氨酸分解模板水凝胶。3D工程组织的尺寸和厚度完全从模板水凝胶转移。通过培养试验、WST-1方法和LIVE/DEAD染色测定来确认在所获得的工程组织中培养的L929细胞的活力。该工程组织是由培养的细胞和由细胞产生的胶原蛋白组成的自站立和高度致密的复合物。这种水凝胶模板的方法可能是有用的，作为一种新的软组织工程技术，以取代合成的聚合物支架的ECM支架从培养的细胞。(c)2007爱思唯尔有限公司保留所有权利。
The three-dimensional (3D)-engineered tissues composed of only cells and extracellular matrices (ECM) were constructed by the hydrogel template approach. The disulfide-crosslinked poly(gamma-glutamic acid) hydrogels were prepared as a template hydrogel. These template hydrogels were easily decomposed under physiological conditions using reductants such as cysteine, glutathione and dithiothreitol by cleavage of disulfide crosslinkage to thiol groups. The decomposed polymers are soluble in cell culture medium. The cleaving of disulfide bond was determined by UV-vis and FT-IR spectroscopies. We successfully prepared the 3D-engineered tissues (thickness/diameter, 2mm/1cm) composed of mouse L929 fibroblast cells and ECM by the decomposition of only the template hydrogel with cysteine after 10 days 3D-cell culture on/in the template hydrogel. The size and thickness of the 3D-engineered tissues was completely transferred from the template hydrogel. The cultured L929 cells viability in the obtained engineered tissues was confirmed by a culture test, WST-1 method and LIVE/DEAD staining assay. The engineered tissue was self-standing and highly dense composite of the cultured cells and collagen produced by the cells. This hydrogel template approach may be useful as a new class of soft-tissue engineering technology to substitute a synthetic polymer scaffold to the ECM scaffold produced from the cultured cells. (c) 2007 Elsevier Ltd. All rights reserved.