Reconstructive UrologyEngineering of Corpus Cavernosum Using Vascular Endothelial Growth Factor-expressing Muscle-derived Stem Cells Seeded on Acellular Corporal Collagen Matrices
Section snippets
Isolation and Expansion of MDSCs
The hind limb gastrocnemius muscles were removed from 2-week-old normal New Zealand white rabbits. MDSCs were obtained and purified by established preplate techniques.13, 19 The proliferation medium was Dulbecco’s modified Eagle’s medium (Gibco, Life Technologies, Carlsband, CA) with 10% fetal bovine serum (Gibco), 10% horse serum (Gibco), 0.5% chick embryo extract (Gibco), and 1% penicillin/streptomycin (Gibco).
Cell Transfection, MDSC Characterization, and VEGF Expression
The human VEGF165 lentiviral gene vector (LV-GFP-VEGF), which contained a green
Transfection of VEGF Gene into MDSCs
After lentiviral transfection, the expression of desmin, Bcl-2, CD34, and CD45, detected using flow cytometry, did not show any significant changes (Fig. 1A). Using fluorescence microscopy, GFP expression in the transfected MDSCs was observed, and transfection efficiency was detected by flow cytometry (Fig. 1B). The exper-iments with MDSCs were performed at a multiplicity of infection of 15. Western blot analysis was used to detected the protein expression of α-SMA in the MDSC, MDSC-vector, and
Comment
Cavernous smooth muscle cells and endothelial cells are the main components of the corpus cavernosum and play an important role in maintaining and regulating the function of the penis. Reconstitution of the corpus cavernosum smooth muscle and sinus endothelium are always the main research components of a TECC. To achieve fully functional tissue-engineered corporal tissue, various approaches have been studied, including a combination of seed cells with biodegradable polymer scaffolds10, 20 or
Conclusion
In the present study, we successfully constructed lentiviral-mediated VEGF-expressing MDSCs and demonstrated their role, as seed cells, in increasing the content of endothelial cells, smooth muscle cells, and capillaries in constructing the TECC. In particular, VEGF-expressing MDSCs greatly increased the capillary density in the engineered corporal tissue compared with simple MDSC. The combination of MDSC seeding with VEGF gene therapy can be considered as a potential new seed cell model to
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Financial Disclosure: The authors declare that they have no relevant financial interests.
Funding Support: This work was supported by the National Natural Science Foundation of China (grant 30672183), the Science and Technology Plan of Guangdong Province (grant 1111220600060), and the Natural Science Foundation of Guangdong Province (grant 8151008901000202).