Decellularization of lung tissue and analysis of its differentiative potential on bone marrow mesenchymal stem cells of rat

Maesomeh Tarahani Nia, Akram Alizadeh, Mohammad Ali Takhshid, Esmaeil Sadroddiny

Abstract


Background: Pulmonary diseases are one of the most common causes of mortality worldwide. In some cases lung transplantation is the only curative treatment. Unfortunately, very few lungs are available for transplantation and also the 5-year survival after lung transplantation is only 50%. Furthermore, transplant recipients require immunosuppressant therapy in that’s time. Currently, production of engineered lung tissue using in vitro stem cells is a promising approach. Extraction of natural ECM or decellularization of lung and application of it in tissue engineering is one of the most important strategies in this regard. Using this technique can preserve the natural characteristics of the ECM and would leads to the removal of the immunogen agents (MHC I, II) and allows reconstruction of graft.

Objective: In this study we have decellularized rat lung and cultured bone marrow stem cells on it to evaluate differentiative potential of it.

Methods: In this study along with extraction of rat lung, its femur and tibia bones were also isolated for extraction of mesenchymal stem cells. Lung tissue was decellularized using SDS detergent. SEM and H&E staining used to assay decellularization. Finally mesenchymal stem cells were seed on the decellularized tissue sections and immunocytochemistry for CC10 and SPD; as markers for differentiation toward lung cells, performed on these cells after 21 days.

Results:In the H&E slides of decellularized tissue there were not any cells. In the electron microscope images of decellularized tissue compared with normal tissue, alveolar structure was well maintained. Also, immunocytochemistry assay showed evidence of differentiation of seeded mesenchymal cells toward lung tissue cells.

Conclusion:In this method decellularization take place without any significant change in tissue structure. Decellularized tissue can induce differentiation of bone marrow stem cells toward lung epithelial cells so natural ECM saved relatively.


Keywords


Decellularization, SDS, Differentiation, CC10, SPD

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References


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