Shrinkage-free preparation of scaffold-free cartilage-like disk-shaped cell sheet using human bone marrow mesenchymal stem cells

被引：20
作者：
Maeda, Satoshi [1 ]
Fujitomo, Takashi [1 ]
Okabe, Takahiro [2 ]
Wakitani, Shigeyuki [2 ]
Takagi, Mutsumi [1 ]
机构：
[1] Hokkaido Univ, Div Biotechnol & Macromol Chem, Grad Sch Engn, Kita Ku, Sapporo, Hokkaido 0608628, Japan
[2] Osaka City Univ, Sch Med, Abeno Ku, Osaka 5458585, Japan
来源：
JOURNAL OF BIOSCIENCE AND BIOENGINEERING | 2011年 / 111卷 / 04期
关键词：
Cartilage regeneration; Mesenchymal stem cells; Shrinkage; Scaffold-free; Chondrogenic differentiation; CHONDROGENIC DIFFERENTIATION; TISSUE; REPAIR; DEFECTS; CULTURE;
D O I：
10.1016/j.jbiosc.2010.11.022
中图分类号：
Q81 [生物工程学（生物技术）]; Q93 [微生物学];
学科分类号：
071005 ; 0836 ; 090102 ; 100705 ;
摘要：
Aiming for the clinical application of cartilage regeneration, a culture method for mesenchymal stem cells (MSCs) derived from human bone marrow to obtain scaffold-free cartilage-like disk-shaped sheet of uniform sizes without the shrinkage was investigated. A disk-shaped cell sheet having the same diameter as that of the membrane without the shrinkage was formed after the cultivation of MSCs (18.6 x 10(5) cells/well) for 3 weeks in a cell culture insert (CCI) containing a flat membrane whose porosity was 12%, while 6.2 and 31.0 x 10(5) MSCs/well, respectively, resulted in the shrinkage of the aggregate and the hole formation in the center part of the sheet. Cell aggregates shrunk also in a 96-well plate and CCIs having lower porosity. The disk-shaped cell sheet showed the comparable thickness (1.2 mm) and sulfated glycosaminoglycan (sGAG) density to those of the pellet formed in a pellet culture. The gene expression levels of aggrecan and type II collagen in the disk-shaped cell sheet were not lower than those in the pellet. In conclusion, the usage of CCI having 12% porosity and 18.6 x 10(5) MSCs/well could avoid the shrinkage from the formation of the scaffold-free cartilage-like disk-shaped cell sheet. (C) 2010, The Society for Biotechnology, Japan. All rights reserved.
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页码：489 / 492
页数：4
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