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. 2014 May 8;4(1):46-51.
eCollection 2014 Jan.

Mesenchymal stem cell response to growth factor treatment and low oxygen tension in 3-dimensional construct environment

Thomas John Sakrison Durant  1 , Nathaniel Dyment  1 , Mary Beth R McCarthy  1 , Mark P Cote  1 , Robert A Arciero  1 , Augustus D Mazzocca  1 , David Rowe  1
Affiliations

Mesenchymal stem cell response to growth factor treatment and low oxygen tension in 3-dimensional construct environment

Thomas John Sakrison Durant et al. Muscles Ligaments Tendons J. .

Abstract

Introduction: cell-based tissue engineering strategies using human mesenchymal stem cells (hMSCs) may help to augment tendon healing. To further investigate the in-vitro behavior of this cell population, we investigated low oxygen culture levels, and growth factor supplementation and their effect on expression of tendon extracellular proteins and cell proliferation.

Materials and methods: bone marrow aspirate (BMA) was harvested during arthroscopic rotator cuff repair. Characterized hMSCs derived from BMA were incorporated into 3-dimensional tissue engineered constructs (TECs). TECs were analyzed by frozen sections with immunohistochemistry for cell density, collagen I and collagen III expression.

Results: growth factor stimulation and low oxygen increased cell density within TECs. Low oxygen and addition of growth factors to culture media demonstrated an increase in collagen I and III expression, both in ambient oxygen conditions and low oxygen conditions.

Conclusion: low oxygen and TGFβ3 demonstrated a positive effect on cell number, and type I and III collagen expression in 3D culture environments.

Keywords: construct; low oxygen; mesenchymal stem cells; tendon; transforming growth factor beta.

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Figures

Figure 1.
Figure 1.
Drawings depicting lateral and top views (left to right) of tissue engineered constructs in silicone gel wells, with suture anchors.
Figure 2.
Figure 2.
Top view of actual tissue engineered construct contracting around suture anchors over time on silicone gel well.
Figure 3.
Figure 3.
Graph depiction of raw pixel counts of type I and III collagen staining within TEC construct.
Figure 4.
Figure 4.
Graph depiction of pixel counts of type I and III collagen staining within TEC construct when normalized to TEC area.
Figure 5.
Figure 5.
Graph depiction of pixel counts of type I and III collagen staining within TEC construct when normalized to TEC area.
See this image and copyright information in PMC

References

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