Differentiation of Blastema Cells in Decellularized Bladder Scaffold in vitro

Biological scaffolds that compose extracellular matrix (ECM) can facilitate the restructuring of a large number of pre-clinical studies in animal tissues and also in clinical applications. As we know interaction between cells and ECM provides suitable three-dimensional structures. The objective of this study is to investigate the effect of induced matrix of New Zealand rabbit bladder on the blastema cells in vitro. In this experimental study, first of all, in order to decellularize, it is necessary to put bladder of New Zealand rabbit in 1% weight/vol solution of sodium dodecyl sulfate (SDS) for 24 hours. Then, to prepare blastema tissue, the pinna of New Zealand rabbit should be punched-hole manually with a special puncher, and after 72 hours a circular blastema is separated with another puncher. After that, the decellularized samples are put in the middle of the blastema ring and transferred to the culture. Then to test, we first use Hematoxylin & Eosin and pick indigo staining. To identify 1 micron samples Toluidine Blue staining is used. After preparing 7 nm sections, some samples are studied by transitional electron microscope on day 15 and 20. Collagen fibers are preserved after decellularizing in matrix of bladder. Most migration of blastema cells occur on days 15 and 20 of culture. Blastema cells are differentiated in different types of cells. Toluidine Blue staining shows desmosome connections between epithelial cells. Glycoproteins that are secreted by the epithelial cells can be indicated in the extracellular matrix (ECM). Bladder scaffold has the ability to induce blastema cells as they differentiate and migrate. This finding is very important since no other growth factor and inducer component has been used.