title: Effect of dynamic seeding methods on the distribution of fibroblasts within human acellular dermis.
abstract: The purpose of this investigation was to compare different dynamic cell seeding methods regarding their seeding efficiency, homogeneity, infiltration depth and proliferation within a human acellular dermis. In addition, the growth behaviour was observed during a 12-day static in vitro culture. The dynamic methods included orbital-shaker seeding and the use of a plate centrifuge with different rotational speeds, combinations of low-pressure for matrix degassing and centrifugal seeding. Scaffolds were incubated for up to 12 days statically. Cell distribution and infiltration depth were analysed histologically at days 0, 4, 8 and 12. Seeding efficiency and cell proliferation were quantified with the MTT-assay at the same time points. Centrifugal seeding with 300g for 5 × 1 min combined with matrix degassing significantly increased the seeding efficiency and homogeneity compared to the other methods. However, following static culture, no cells were detectable after 4 days in the inner matrix zones. Furthermore, none of the degassing+centrifugation groups reached a significantly higher proliferation at day 8 compared to the reference. The use of a single dynamic method resulted in an inefficient cell seeding. We archived the highest seeding efficiency, homogeneity and infiltration depth using a combination of degassing+centrifugation at 300g for 5 × 1 min.
authors: Division of Surgical Oncology and Thoracic Surgery, Department of Surgery, University Medical Centre Mannheim, University of Heidelberg, Theodor- Kutzer- Ufer 1-3, 68167, Mannheim, Germany. m.vitacolonna@gmail.com.
authors: Mario
authors: M
authors: Vitacolonna
authors: Institute of Pathology, University Medical Centre Mannheim, University of Heidelberg, 68167, Mannheim, Germany. djeda.belharazem@medma.uni-heidelberg.de.
authors: Djeda
authors: D
authors: Belharazem
authors: Division of Surgical Oncology and Thoracic Surgery, Department of Surgery, University Medical Centre Mannheim, University of Heidelberg, Theodor- Kutzer- Ufer 1-3, 68167, Mannheim, Germany. peter.hohenberger@medma.uni-heidelberg.de.
authors: Peter
authors: P
authors: Hohenberger
authors: Division of Surgical Oncology and Thoracic Surgery, Department of Surgery, University Medical Centre Mannheim, University of Heidelberg, Theodor- Kutzer- Ufer 1-3, 68167, Mannheim, Germany. eric.roessner@medma.uni-heidelberg.de.
authors: Eric D
authors: ED
authors: Roessner
mesh_terms: D062088:Acellular Dermis; D061249:Batch Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D002474:Cell-Free System / Q000737:chemistry; D002478:Cells, Cultured; D005347:Fibroblasts / Q000166:cytology* / Q000502:physiology* / Q000637:transplantation; D006801:Humans; D054457:Tissue Scaffolds
keywords: Cell seeding; Comparison of dynamic seeding methods; Dynamic cell seeding methods; Human acellular dermis; Scaffold seeding; Tissue-Engineering