Author/Editor		Kregar-Velikonja, Nevenka
Title		Diferenciacija in tkivno specifično izražanje genov v kulturi humanega hialinega hrustanca
Translated title		Differentiation and tissue specific gene expression in cultures of human hialine cartilage
Type		monografija
Place		Ljubljana
Publisher		Medicinska fakulteta
Publication year		2002
Volume		str. 126
Language		slo
Abstract		Developments in tissue engineering have led to the routine treatment of cartilage lesions in knee joint with autologous chondrocytes, cultured in vitro. This approach is currently being developed towards the engineering of 3-D cartilage constructs and the use of mesenchymal stem cells. We studied two cell types that can be used for engineering of cartilaginous tissue: chondrocytes (HC), isolated from human articular cartilage, and bone marrow derived mesenchymal cells (MSC), that were proven to have multipotent character. The ffow cytometry analysis using cell membrane markers CD90, CD105 and CD106 has been developed for the determination of both cell types. We tested the influence of several growth and differentiation factors on cell proliferation rates and the expression of cartilage specific genes. Platelet derived growth factor was found to be the most efficient promoter of HC and MSC proliferation, while TGF(3 was the most efficient factor in modifying specific gene expression. The both cell types were used for the preparation of 3D cartilaginous constructs in vitro. We used serologic, histologic and molecular biology methods for the assessment of cartilage phenotype. We were able to detect increased synthesis of collagen type II and aggrecan in the presence of HG I and HG II media. In parallel the changes in expression of Sox 9, Cep 68 and SH2 genes were also detected. The chondrogenic differentiation was more abundant in the high density cell cultures, as compared to the cultures, grown on the three dimensional collagen scaffolds. Mesenchymal stem cells are more suitable for preparation of long term 3D cultures. They also form homogenous layer of tissue with a thickness up to 0,2 mm, when seeded on polycarbonate membrane support. (Abstract truncated at 2000 characters).
Descriptors		GENE EXPRESSION HYALIN CARTILAGE, ARTICULAR CELLS, CULTURED HEMATOPOIETIC STEM CELLS FLOW CYTOMETRY PHENOTYPE CELL DIFFERENTIATION IMMUNOENZYME TECHNIQUES POLYMERASE CHAIN REACTION MICROSCOPY, CONFOCAL