title: Comparison of analytical methods for quantitation of isolated porcine hepatocyte yields.
abstract: As cell-based therapies receive approval for clinical evaluation and use, the development of reliable methods to quantify cell number and control the dose of therapy delivered is becoming increasingly important. An example is the determination of the number and volume of primary porcine hepatocytes used in an extracorporeal treatment for patients with liver disease. Conventional cell counting using optical microscopy was compared against two alternate methods to quantify isolated porcine hepatocytes: (1) automated cell counting using a commercially available particle characterization instrument, and (2) quantitation by cell mass. Methods were compared based on accuracy, precision, specificity, linear range, and ruggedness. The automated method delivered substantially improved accuracy, precision, and ruggedness when compared to the conventional optical method. It also provided valuable information about the size distribution of cell preparations, which often contained clumps of cells, and showed that processing steps such as cryopreservation can alter the size characteristics of a cell population. The automated method was also faster, and was well suited to use in a commercial manufacturing process. The mass-based method was simple and inexpensive, but suffered from nonlinearity at low cell concentrations. Automated cell quantitation using a commercially available particle characterization instrument proved to be the preferred method for obtaining accurate and consistent porcine hepatocyte counts in a timely manner.
authors.affiliation: Department of Biomedical Engineering, Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, GA, 30332-0535, USA.
authors.forename: J P
authors.initials: JP
authors.lastname: Stegemann
authors.forename: S
authors.initials: S
authors.lastname: Raina
authors.forename: D T
authors.initials: DT
authors.lastname: Nicholson
authors.forename: P
authors.initials: P
authors.lastname: Jimenez
authors.forename: L
authors.initials: L
authors.lastname: Shah
authors.forename: S
authors.initials: S
authors.lastname: Cain
authors.forename: B
authors.initials: B
authors.lastname: Chandler
authors.forename: Z
authors.initials: Z
authors.lastname: Pitkin
authors.forename: C
authors.initials: C
authors.lastname: Mullon
authors.forename: L
authors.initials: L
authors.lastname: Custer
mesh_terms: D000818:Animals; D002452:Cell Count / Q000379:methods*; D017690:Cell Transplantation; D008099:Liver / Q000166:cytology*; D016031:Liver Transplantation; D013552:Swine