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Bioreactor conditioning of valve scaffolds seeded internally with adult stem cells

  • Original Article
  • Tissue Engineering
  • Published:
Tissue Engineering and Regenerative Medicine Aims and scope

Abstract

The goal of this study was to test the hypothesis that stem cells, as a response to valve-specific extracellular matrix “niches” and mechanical stimuli, would differentiate into valvular interstitial cells (VICs). Porcine aortic root scaffolds were prepared by decellularization. After verifying that roots exhibited adequate hemodynamics in vitro, we seeded human adipose-derived stem cells (hADSCs) within the interstitium of the cusps and subjected the valves to in vitro pulsatile bioreactor testing in pulmonary pressures and flow conditions. As controls we incubated cell-seeded valves in a rotator device which allowed fluid to flow through the valves ensuring gas and nutrient exchange without subjecting the cusps to significant stress. After 24 days of conditioning, valves were analyzed for cell phenotype using immunohistochemistry for vimentin, alpha-smooth muscle cell actin (SMA) and prolyl-hydroxylase (PHA). Fresh native valves were used as immunohistochemistry controls. Analysis of bioreactor-conditioned valves showed that almost all seeded cells had died and large islands of cell debris were found within each cusp. Remnants of cells were positive for vimentin. Cell seeded controls, which were only rotated slowly to ensure gas and nutrient exchange, maintained about 50% of cells alive; these cells were positive for vimentin and negative for alpha-SMA and PHA, similar to native VICs. These results highlight for the first time the extreme vulnerability of hADSCs to valve-specific mechanical forces and also suggest that careful, progressive mechanical adaptation to valve-specific forces might encourage stem cell differentiation towards the VIC phenotype.

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Authors and Affiliations

  1. Biocompatibility and Tissue Regeneration Laboratories, Department of Bioengineering, Clemson University, 304 Rhodes Annex, Clemson, SC, 29634, USA

    Allison Kennamer, Leslie Sierad, Richard Pascal, Nicholas Rierson, Christopher Albers & Dan Simionescu

  2. Tissue Engineering and Regenerative Medicine Laboratory, Department of Anatomy, University of Medicine and Pharmacy, Targu Mures, Romania

    Marius Harpa, Ovidiu Cotoi, Lucian Harceaga, Peter Olah, Preda Terezia & Dan Simionescu

  3. Cardiovascular Tissue Engineering and Regenerative Medicine Laboratory, Department of Bioengineering, Clemson University, Clemson, SC, USA

    Agneta Simionescu

Authors
  1. Allison Kennamer
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  2. Leslie Sierad
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  3. Richard Pascal
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  4. Nicholas Rierson
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  5. Christopher Albers
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  6. Marius Harpa
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  7. Ovidiu Cotoi
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  8. Lucian Harceaga
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  9. Peter Olah
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  10. Preda Terezia
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  11. Agneta Simionescu
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  12. Dan Simionescu
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Correspondence to Dan Simionescu.

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Kennamer, A., Sierad, L., Pascal, R. et al. Bioreactor conditioning of valve scaffolds seeded internally with adult stem cells. Tissue Eng Regen Med 13, 507–515 (2016). https://doi.org/10.1007/s13770-016-9114-1

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  • DOI: https://doi.org/10.1007/s13770-016-9114-1

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