Liver tissue engineering within alginate scaffolds: Effects of cell-seeding density on hepatocyte viability, morphology, and function
Liver tissue engineering within alginate scaffolds: Effects of cell-seeding density on hepatocyte viability, morphology, and function
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DOI：
10.1089/107632703768247430
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发表时间：
2003-08-01
影响因子：
--
通讯作者：
Cohen, S
中科院分区：
文献类型：
--
作者：
Dvir-Ginzberg, M;Gamlieli-Bonshtein, I;Cohen, S
三维生物材料的组织工程是一种很有前途的方法，用于开发肝组织来替代功能衰竭的肝脏。在这里，我们解决细胞接种和分布在多孔藻酸盐支架，这代表了一种新型的多孔生物材料的组织工程。藻酸盐支架的亲水性以及其孔结构和互连性使得能够将肝细胞有效地接种到支架中，即，取决于接种方法，初始细胞的70-90%。在接种期间利用离心力增强了多孔支架中的细胞分布，因此能够接种浓缩的细胞悬浮液（>1 X 10（7）个细胞/mL）。MTT法检测支架中细胞密度对肝细胞活力的影响。在细胞密度为0.28 × 10（6）个细胞/cm（3）支架时，活肝细胞的数量在7天内下降至初始值的33%，而在密度为5.7 × 10（6）个细胞/cm（3）支架或更高的培养物中，细胞保持较高的活力，同时形成连接球状体的网络。在高密度的细胞结构，肝细胞的功能，如白蛋白和尿素的分泌，解毒（细胞色素P-450和II相结合酶的活动），保持在7天的文化高。总的来说，本研究的结果突出了细胞密度对三维肝细胞构建体的肝细胞功能的重要性以及藻酸盐基质作为支架的优势。
Tissue engineering with three-dimensional biomaterials represents a promising approach for developing hepatic tissue to replace the function of a failing liver. Herein, we address cell seeding and distribution within porous alginate scaffolds, which represent a new type of porous biomaterial for tissue engineering. The hydrophilic nature of the alginate scaffold as well as its pore structure and interconnectivity enabled the efficient seeding of hepatocytes into the scaffolds, that is, 70-90% of the initial cells depending on the seeding method. Utilization of centrifugal force during seeding enhanced cell distribution in the porous scaffolds, consequently enabling the seeding of concentrated cell suspensions (>1 X 10(7) cells/mL). Cell density in scaffolds affected hepatocyte viability as judged by MTT assay. At a cell density of 0.28 X 10(6) cells/cm(3) scaffold, the number of viable hepatocytes decreased to 33% of its initial value within 7 days, whereas at the denser cultures, 5.7 X 10(6) cells/cm(3) scaffold and higher, the cells maintained higher viability while forming a network of connecting spheroids. In the high-density cellular constructs, hepatocellular functions such as albumin and urea secretion, and detoxification (cytochrome P-450 and phase II conjugating enzyme activities), remained high during the 7-day culture. Collectively, the results of the present study highlight the importance of cell density on the hepatocellular functions of three-dimensional hepatocyte constructs as well as the advantages of alginate matrices as scaffoldings.