Comparative study of decelluralized extracellular matrix from porcine derived tissues as a substrate for in-vitro mouse mesenchymal stem cell culture



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The main goal of this research is to create a 3-D natural scaffold for the Mouse Mesenchymal stem cells (MSCs) to carry out in-vitro growth without losing its physiological and morphological characteristics. MSCs are multipotent adult stem cells that can develop into several cell types belonging to bone, skeletal and fat tissues. These cells require extracellular matrix (ECM) for their growth. ECM is a critical environmental factor for cells to maintain normal function. It provides structural and biochemical support to them. Without ECM substrates, in-vivo and in-vitro stem cell research is of limited use. Furthermore, therapeutic applications require large numbers of MSCs. Because of MSCs’s biological importance and diverse role, the ECM has been the focal point of increasing interest in the field of regenerative medicine and stem cell research. Therefore, tissue culture plates have been developed providing ECM components to maintain an artificial environment for the cells to grow. However, this approach has limited success since it provides a flat 2-D growth surface and hence the standard culturing techniques have proven insufficient for this purpose. In addition, the tissue culture plates are expensive. This research is to prepare a complete ECM comprised of all the components required to provide a natural environment for the growth of healthy cells with intact tissues. The context discussed indicates that minimally altered decellularized porcine tissues-derived ECM can provide effective substrates for MSCs. In the present study it was found that bone marrow (BM), adipose and dermis smeared cultivation plates support in-vitro cultivation of MSCs while maintaining homogenous, physiological and stable cell population. Characterization and analyses of BM, adipose and dermis was carried out by performing immunohistochemistry and other staining techniques which demonstrate the presence of major ECM proteins and glycosaminoglycans (GAGs) in the substrates. The results of this research will establish a porcine derived ECM substrate with application potential in high fidelity cultivation techniques for stem cells. On the basis of our results, we suggest that decellularized ECM has a significant impact on tissue reconstruction and regenerative medicine.