Department of Chemical Engineering and Biochemical Engineering, Zhejiang University, 38 Zheda Road, 310027, Hangzhou, Zhejiang, Peoples Republic of China.
Cytotechnology. 2009 Jul;60(1-3):133-41. doi: 10.1007/s10616-009-9228-6. Epub 2009 Sep 27.
Gel entrapment culture of primary mammalian cells within collagen gel is one important configuration for construction of bioartificial organ as well as in vitro model for predicting drug situation in vivo. Gel contraction in entrapment culture, resulting from cell-mediated reorganization of the extracellular matrix, was commonly used to estimate cell viability. However, the exact influence of gel contraction on cell activities has rarely been addressed. This paper investigated the gel contraction under varying culture conditions and its effect on the activities of rat hepatocyte entrapped in collagen gel within hollow fibers. The hepatocyte activities were reflected by cell viability together with liver-specific functions on urea secretion and cytochrome P450 2E1. Unexpectedly, no gel contraction occurred during gel entrapment culture of hepatocyte under a high collagen concentration, but hepatocytes still maintained cell viability and liver-specific functions at a similar level to the other cultures with normal gel contraction. It seems that cell activities are unassociated with gel contraction. Alternatively, the mass transfer resistance induced by the combined effect of collagen concentration, gel contraction and cell density could be a side effect to reduce cell activities. The findings with gel entrapment culture of hepatocytes would be also informative for the other cell culture targeting pathological studies and tissue engineering.
胶内原代哺乳动物细胞的包埋培养是构建生物人工器官和预测体内药物情况的体外模型的重要方法之一。胶内包埋培养中的凝胶收缩是由细胞介导的细胞外基质的重新组织引起的，常用于估计细胞活力。然而，凝胶收缩对细胞活性的确切影响很少被涉及。本文研究了不同培养条件下的凝胶收缩及其对胶原凝胶中空纤维内包埋大鼠肝细胞活性的影响。细胞活力和尿素分泌、细胞色素 P4502E1 等肝脏特异性功能反映了肝细胞的活性。出乎意料的是，在高浓度胶原的情况下，肝细胞在胶内包埋培养过程中并没有发生凝胶收缩，但仍保持着类似的细胞活力和肝脏特异性功能，与其他正常凝胶收缩的培养相比。似乎细胞活性与凝胶收缩无关。或者，胶原浓度、凝胶收缩和细胞密度的综合作用引起的传质阻力可能是降低细胞活性的副作用。这些关于肝细胞胶内包埋培养的发现对于其他针对病理研究和组织工程的细胞培养也具有重要意义。
