MTS colorimetric assay in combination with a live-dead assay for testing encapsulated L929 fibroblasts in alginate poly-L-lysine microcapsules in vitro.
Biomaterials such as applied in microcapsules may have harmful effects on encapsulated cells. Up to now, there are no adequate assays available for testing the function and viability of cells in capsules. Therefore, we investigated whether the combination of MTS proliferation assay and live-dead viability assay is suitable for testing microencapsulated L929 fibroblasts in long-time culture. Proliferation of L929 cells was shown by a significant increase of formazan absorbance within the first 3 weeks (Day 0: 0.132 +/- 0.047; Day 7: 0.404 +/- 0.101; Day 14: 0.728 +/- 0.239; Day 21: 0.877 +/- 0.224) followed by stagnation and decrease thereafter. This was confirmed by an increasing proportion of dead cells measured by the live-dead assay. Thus, proliferation of encapsulated L929 can be reliably investigated by the MTS assay. In combination with life-dead assays, the proliferation can be correlated to the survival rate of the encapsulated cells.
Bünger
C M
CM
Department of Surgery, University of Rostock, Rostock, Germany. carsten.buenger@medizin.uni-rostock.de
Jahnke
A
A
Stange
J
J
de Vos
P
P
Hopt
U T
UT
D000464:Alginates; D000818:Animals; D001672:Biocompatible Materials; D002455:Cell Division; D002478:Cells, Cultured; D003124:Colorimetry; D004396:Coloring Agents; D005347:Fibroblasts / Q000166:cytology; D005562:Formazans; D006801:Humans; D008567:Membranes, Artificial; D051379:Mice; D011107:Polylysine / Q000031:analogs & derivatives*; D014343:Trypan Blue
D000464:Alginates; D001672:Biocompatible Materials; D004396:Coloring Agents; D005562:Formazans; D008567:Membranes, Artificial; C051546:alginate-polylysine-alginate; D011107:Polylysine; C059088:1,5-bis(2-methoxy-4-nitro-5-sulfophenyl)-3-((phenylamino)carbonyl)formazan; D014343:Trypan Blue