Determination of the decay rate constant for hepatocytes immobilized in alginate microcapsules.
Primary mouse hepatocytes (between 10-250 cells per capsule) were immobilized within 1.0% w/v alginate microbeads. The textural properties of the alginate matrix were characterized and a full protocol based upon the measurement of the initial rate of Resazurin reduction was studied and standardized. Using this method, the decay rate constant (K(d) = 0.45 +/- 0.01 days(-1)) and the time in which the cell viability decreases in half (VI(50) = 37 +/- 0.7 h) have been measured. The method was compared with the analysis of cell vitality using Calcein A/M and Ethidium Homodimer I. Differences between the two methods were found in the viability profile due to the significant presence of double stained cells along the culture time. According to the author's knowledge, this is the first report of a systematic study and determination of the K(d) value for immobilized hepatocytes, incorporating a wide range of cell concentrations within the alginate matrix.
Leal-Egaña
Aldo
A
Department of Cell Technology and Applied Stem Cell Biology, Biotechnology and Biomedicine Centre (BBZ), University of Leipzig, Leipzig, Germany. leal@rz.uni-leipzig.de
Díaz-Cuenca
Aránzazu
A
Bader
Augustinus
A
D000464:Alginates / Q000737:chemistry; D000818:Animals; D002214:Capsules / Q000737:chemistry; D002470:Cell Survival; D002478:Cells, Cultured; D018914:Cells, Immobilized / Q000166:cytology; D005260:Female; D020723:Glucuronic Acid / Q000737:chemistry; D022781:Hepatocytes / Q000166:cytology*; D006603:Hexuronic Acids / Q000737:chemistry; D007202:Indicators and Reagents / Q000737:chemistry; D051379:Mice; D010078:Oxazines / Q000737:chemistry; D010084:Oxidation-Reduction; D016062:Porosity; D014966:Xanthenes / Q000737:chemistry
D000464:Alginates; D002214:Capsules; D006603:Hexuronic Acids; D007202:Indicators and Reagents; D010078:Oxazines; D014966:Xanthenes; C005843:resazurin; D020723:Glucuronic Acid