title: Static image analysis as new approach for the characterization of tumor cell lysate used in dendritic cell vaccine preparation.
abstract: BACKGROUND
  Safety is an important consideration for the clinical application of dendritic cells (DC) loaded with autologous tumor lysate (TL). Thus, avitalization of TL from living autologous tumor tissue has to be guaranteed.
  
  
  METHODS
  Composition of TL was investigated by static image analysis (SIA) with the Morphologi G3 device, which simultaneously measures size and shape of up to 100,000 particles within one sample run. This approach was compared with sample characterization by high-resolution automated cell counting, trypan blue staining, and ATP quantification.
  
  
  RESULTS
  Using SIA, we only detected fragmented, non-cellular structures in completely avitalized TL, indicating complete destruction of living cells. Analysis of particle size distribution by SIA as well as CASY cell counter showed that 95% of particles had a diameter of <10 µm as a sign of cell fragmentation. Complete avitalization of TL was confirmed with trypan blue staining and ATP analysis.
  
  
  CONCLUSIONS
  Regarding generation of DC vaccines, the proof of avitality of TL from living tumor tissue can clearly be achieved by SIA alone or in combination with standard assays. Our data show that SIA is a highly precise method for TL characterization. The SIA device complies with FDA regulation and, therefore, might be suitable for characterization of cellular therapy medicinal products.
authors.affiliation: Institute of Clinical Hemostaseology and Transfusion Medicine, Saarland University Medical Center, Homburg, Germany.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Müller
authors.affiliation: Institute of Clinical Hemostaseology and Transfusion Medicine, Saarland University Medical Center, Homburg, Germany.
authors.forename: Dorothee
authors.initials: D
authors.lastname: Hartmann
authors.affiliation: Department of Neurosurgery, Saarland University Medical Center, Homburg, Germany.
authors.forename: Joachim
authors.initials: J
authors.lastname: Oertel
authors.affiliation: Applied Pharmacy, University of Applied Sciences Kaiserslautern, Pirmasens, Germany.
authors.forename: Cornelia M
authors.initials: CM
authors.lastname: Keck
authors.affiliation: Institute of Clinical Hemostaseology and Transfusion Medicine, Saarland University Medical Center, Homburg, Germany.
authors.forename: Hermann
authors.initials: H
authors.lastname: Eichler
keywords: Autologous tumor lysate; Avitality; Bioinactivity; DC vaccine