The use of 3D culture of hanging drop mesenchymal stem cells in collagen scaffolds for tissue-engineered repair of cartilage

Introduction The aim of regenerative medicine is to revive injured tissues by developing efficient cell, tissue, and organ replacements to repair or increase biological function in damaged tissues. We hypothesized that the 3D culture of mesenchymal stem cells (MSCs) in collagen scaffold (collagen rat tail) can improve stem cells differentiation into cartilage. With regard to these considerations, the current study aimed to reconstruct a culture environment for cartilage tissue engineering.Methods The human MSCs derived from Wharton‘s jelly were cultured in completed Dulbecco s Modified Eagle s medium (DMEM) in primary culture. Then the cell were characterized as MSCs by flow-cytometry. The cell were cultured in hanging drop. The collagen scaffold was made and lyophilized. Then, the 3D culture hanging drop cell put on collagen scaffolds during 2 weeks. Glycosaminoglycans (GAGs) was identified in extracellular matrix of MSCs by safarin o and alcian blue staining. All the scaffolds were evaluated by scanning electron microscope (SEM) and Fourier Transforms Infrared Spectrometry (FTIR). Scaffold porosity was also determined. The differentiation of MSCs into chondrocyte was also compared with control group.ResultsFTIR and SEM analysis identified that this scaffold are highly porous with interconnection. This can provide an ideal matrix which is suitable for cell growth and chondrocyte differentiation. The cells were differentiate to chondrocyte in this 3d porous scaffolds that can be useful in tissue engineering.ConclusionsThe cell cultured in a new method showed that a different physical and biological property is useful in cartilage tissue engineering applications

کلیدواژه ها:

Scaffold ، Mesenchymal stem cells ، 3D culture ، Hanging drop