Application of colorimetric assays to assess viability, growth and metabolism of hydrogel-encapsulated cells.
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.
Khattak
Sarwat F
SF
Department of Chemical Engineering, University of Massachusetts, 686 North Pleasant Street, Amherst, MA 01003, USA.
Spatara
Michelle
M
Roberts
Louis
L
Roberts
Susan C
SC
D000464:Alginates; D000818:Animals; D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D003124:Colorimetry / Q000379:methods*; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D007770:L-Lactate Dehydrogenase / Q000032:analysis*; D051379:Mice; D051381:Rats; D013778:Tetrazolium Salts / Q000378:metabolism*; D013844:Thiazoles / Q000378:metabolism*; D023822:Tissue Engineering
D000464:Alginates; D006603:Hexuronic Acids; D013778:Tetrazolium Salts; D013844:Thiazoles; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D020723:Glucuronic Acid; D007770:L-Lactate Dehydrogenase; C022616:thiazolyl blue