Tissue-engineered trachea from sheep marrow stromal cells with transforming growth factor β2 released from biodegradable microspheres in a nude rat recipient

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Tissue-engineered trachea from sheep marrow stromal cells with transforming growth factor β2 released from biodegradable microspheres in a nude rat recipient Koji Kojima, MD, PhD, Ronald A Ignotz, PhD, Toshihiro Kushibiki, MS, Kevin W Tinsley, MA, Yasuhiko Tabata, PhD, DMedSci, DPharm, Charles A Vacanti, MD The Journal of Thoracic and Cardiovascular Surgery Volume 128, Issue 1, Pages 147-153 (July 2004) DOI: 10.1016/j.jtcvs.2004.02.038

Figure 1 Phase-contrast photomicrograph at 200× of sheep MSCs in monolayer culture. A, Cells attaching and colonizing surface of culture flask at 4 days. B, Cells continued to proliferate, and after 14 days initial culture became confluent. The Journal of Thoracic and Cardiovascular Surgery 2004 128, 147-153DOI: (10.1016/j.jtcvs.2004.02.038)

Figure 2 A, Cells attaching to PGA on day 0. B, Growth of cells and matrix on day 7. C, Electron microscopy shows cells firmly attached to PGA by day 4. The Journal of Thoracic and Cardiovascular Surgery 2004 128, 147-153DOI: (10.1016/j.jtcvs.2004.02.038)

Figure 3 A, Cell-seeded matrix was placed in grooves of helical templates fabricated with silicone mold-making kit. B, Constructs coated with gelatin microspheres incorporating TGF-β2 were implanted into subcutaneous pockets on nude rats. The Journal of Thoracic and Cardiovascular Surgery 2004 128, 147-153DOI: (10.1016/j.jtcvs.2004.02.038)

Figure 4 TET from nude rat at 6 weeks after implantation. The Journal of Thoracic and Cardiovascular Surgery 2004 128, 147-153DOI: (10.1016/j.jtcvs.2004.02.038)

Figure 5 Hematoxylin and eosin staining (magnification 100×) of TET (A) and native trachea (B). Safranin-O staining was deeply positive, indicative of abundant proteoglycan production, in both TET (C) and native trachea (D). Staining for type collagen II was strongly positive in extensive extracellular matrix of engineered (E) and native (F) sections. The Journal of Thoracic and Cardiovascular Surgery 2004 128, 147-153DOI: (10.1016/j.jtcvs.2004.02.038)

Figure 6 A, Glycosaminoglycan (GAG) contents of native trachea and TET. B, Hydroxyproline contents of native trachea and TET. Bar heights represent mean of 6 observations; error bars represent SD. Data were not statistically different according to 1-way analysis of variance. The Journal of Thoracic and Cardiovascular Surgery 2004 128, 147-153DOI: (10.1016/j.jtcvs.2004.02.038)