Khattak Sarwat F, Spatara Michelle, Roberts Louis, Roberts Susan C
Department of Chemical Engineering, University of Massachusetts, 686 North Pleasant Street, Amherst, MA 01003, USA.
Biotechnol Lett. 2006 Sep;28(17):1361-70. doi: 10.1007/s10529-006-9104-9. Epub 2006 Jul 4.
The applicability of the colorimetric 3-(4,5-dimethylthiozol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) assays to measure cell growth and viability in hydrogel encapsulation systems was investigated using HepG2 liver cells encapsulated in alginate matrices. The MTT assay was effective in measuring viable cell density in alginate-encapsulated cell systems, demonstrating less variance and higher throughput capability than hemocytometry. The LDH assay was effective in measuring dead cell density in monolayer cultures and in alginate-encapsulated cells simply by measuring the LDH concentration secreted into the medium. Further validation of these assays was shown in two additional cell lines (rat muscle and mouse embryonic fibroblasts). The MTT and LDH assays are particularly significant in the rapid evaluation of in vitro cell encapsulation device design.
使用包封在藻酸盐基质中的HepG2肝细胞,研究了比色法3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)和乳酸脱氢酶(LDH)测定法在水凝胶包封系统中测量细胞生长和活力的适用性。MTT测定法在测量藻酸盐包封细胞系统中的活细胞密度方面是有效的,与血细胞计数法相比,显示出更小的差异和更高的通量能力。LDH测定法通过简单地测量分泌到培养基中的LDH浓度,在测量单层培养物和藻酸盐包封细胞中的死细胞密度方面是有效的。在另外两种细胞系(大鼠肌肉细胞和小鼠胚胎成纤维细胞)中也显示了这些测定法的进一步验证。MTT和LDH测定法在体外细胞包封装置设计的快速评估中特别重要。