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Effect of dynamic seeding methods on the distribution of fibroblasts within human acellular dermis

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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.

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Acknowledgments

The authors thank Dr. Mark D. Smith and Dr. Jan C. Brune from the German Institute for Cell and Tissue Replacement (DIZG) for advice relating to use of Epiflex® and for assistance with data analysis and editing of the final manuscript. The authors also thank Dr. Lothar Pilz from the Medical Faculty Mannheim for his assistance with the statistical analyses.

Conflict of interest

Dr. Mario Vitacolonna, Dr. Djeda Belharazem, Prof. Dr. Peter Hohenberger and Dr. Eric Roessner declare that they have no conflict of interest.

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Authors and Affiliations

  1. 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

    Mario Vitacolonna, Peter Hohenberger & Eric D. Roessner

  2. Institute of Pathology, University Medical Centre Mannheim, University of Heidelberg, 68167, Mannheim, Germany

    Djeda Belharazem

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  1. Mario Vitacolonna
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  2. Djeda Belharazem
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  3. Peter Hohenberger
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  4. Eric D. Roessner
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Correspondence to Mario Vitacolonna.

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Vitacolonna, M., Belharazem, D., Hohenberger, P. et al. Effect of dynamic seeding methods on the distribution of fibroblasts within human acellular dermis. Cell Tissue Bank 16, 605–614 (2015). https://doi.org/10.1007/s10561-015-9508-7

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  • DOI: https://doi.org/10.1007/s10561-015-9508-7

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