Xeno-free and shrinkage-free preparation of scaffold-free cartilage-like disc-shaped cell sheet using human bone marrow mesenchymal stem cells(CELL AND TISSUE ENGINEERING)
スポンサーリンク
概要
- 論文の詳細を見る
Aiming for the clinical application of cartilage regeneration, the xeno-free cultivation method to obtain a scaffold-free cartilage-like disc-shaped cell sheet using mesenchymal stem cells (MSCs) derived from human bone marrow without the shrinkage of the sheet was investigated. MSCs were inoculated into Cell Culture Insert (0.3 cm^2, pore size; 0.4 μm, pore density; 1.0 × 10^8/cm^2) using serum-free chondrogenic differentiation medium containing TGF-β3, IGF-1 and dexamethasone or other modified media, and cultured at 37℃ in 5% CO_2 for 3 weeks. Sheet thickness, cartilage specific genes expression, ECM accumulation were determined, and the sections of sheets were stained with alcian blue. A novel mixed medium consisting of a growth medium (10% FCS) with a serum-free chondrogenic differentiation medium could prevent the shrinkage of the sheet and produced a disc-shaped cell sheet. The depth of the sheet was approximately 0.7 mm and the gene expression levels were higher than those in cells in normal human cartilage. The use of human serum instead of FCS did not cause shrinkage and did not decrease the accumulation levels of sGAG and type 2 collagen in the sheet. The cultivation of MSCs grown with completely xeno-free materials using the mixed medium containing human serum in a cell culture insert showed a sheet depth of 1.0 mm and gene expression levels higher than those in normal cartilage. The scaffold-free and xeno-free cartilage-like cell sheet was successfully formed without shrinkage using human bone marrow MSCs and the chondrogenic differentiation medium containing human serum.
著者
-
Takagi Mutsumi
Division Of Biotechnology And Macromolecular Chemistry Graduate School Of Engineering Hokkaido Unive
-
Sato Yasushi
Division Of Biotechnology And Macromolecular Chemistry Graduate School Of Engineering Hokkaido Unive
-
Wakitani Shigeyuki
School of Health and Sports Sciences, Mukogawa Women's University
-
Sato Yasushi
Division of Biotechnology and Macromolecular Chemistry, Graduate School of Engineering, Hokkaido University
関連論文
- Noninvasive discrimination of human normal cells and malignant tumor cells by phase-shifting laser microscopy(CELL AND TISSUE ENGINEERING)
- Effect of Monosaccharides Composing Glycosaminoglycans on Type 2 Collagen Accumulation in a Three-Dimensional Culture of Chondrocytes(CELL AND TISSUE ENGINEERING)
- A mass separation of chondrocytes from cartilage tissue utilizing an automatic crushing device(CELL AND TISSUE ENGINEERING)
- Fetal calf serum-free suspension culture of Chinese hamster ovary cells employing fish serum(CELL AND TISSUE ENGINEERING)
- Influence of Fetal Calf Serum on Differentiation of Mesenchymal Stem Cells to Chondrocytes during Expansion(CELL AND TISSUE ENGINEERING)
- Combination of cytomegalovirus enhancer with human cellular promoters for gene-induced chondrogenesis of human bone marrow mesenchymal stem cells(CELL AND TISSUE ENGINEERING)
- Cell Processing Engineering for Ex-Vivo Expansion of Hematopoietic Cells
- Effect of Static Pressure on Intracellular pH of Adhesive Chinese Hamster Ovary Cells(CELL AND TISSUE ENGINEERING)
- Development of hybrid artificial lung with gene transfected biological cells
- High inoculation cell density could accelerate the differentiation of human bone marrow mesenchymal stem cells to chondrocyte cells.
- Aortic Root Replacement with a Mechanical Valve and Prosthetic Conduit for the Complicated Degeneration of the Ascending Aorta Resulting from Infective Endocarditis
- Ex vivo cartilage defect model for the evaluation of cartilage regeneration using mesenchymal stem cells(CELL AND TISSUE ENGINEERING)
- Quantitative correlation between production rate of melanoma inhibitory activity and aggrecan gene expression level during differentiation from mesenchymal stem cells to chondrocytes and redifferentiation of chondrocytes(CELL AND TISSUE ENGINEERING)
- Shrinkage-free preparation of scaffold-free cartilage-like disk-shaped cell sheet using human bone marrow mesenchymal stem cells(CELL AND TISSUE ENGINEERING)
- Aortic Valve Replacement in Elderly Aortic Stenosis Patients
- Time-lapse analysis of laser phase shift for noninvasive discrimination of human normal cells and malignant tumor cells(CELL AND TISSUE ENGINEERING)
- Correlation between actin content and laser phase shift of adhesive normal and malignant prostate epithelial cells(CELL AND TISSUE ENGINEERING)
- The Influence of Preexistent Renal Failure on the Prognosis Following Ruptured Abdominal Aortic Aneurysms.
- Xeno-free and shrinkage-free preparation of scaffold-free cartilage-like disc-shaped cell sheet using human bone marrow mesenchymal stem cells(CELL AND TISSUE ENGINEERING)