Three-dimensional microenvironments retain chondrocyte phenotypes during proliferation culture.
Three-dimensional microenvironments retain chondrocyte phenotypes during proliferation culture.
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DOI：
10.1089/ten.2006.0322
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发表时间：
2007-07
影响因子：
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通讯作者：
Tsuguharu Takahashi;T. Ogasawara;Y. Asawa;Y. Mori;E. Uchinuma;T. Takato;K. Hoshi
中科院分区：
文献类型：
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作者：
Tsuguharu Takahashi;T. Ogasawara;Y. Asawa;Y. Mori;E. Uchinuma;T. Takato;K. Hoshi
虽然自体软骨细胞移植已经在临床上使用，软骨细胞去分化是一个问题，在增殖培养。我们尝试在软骨细胞增殖下进行三维（3D）胶原凝胶培养，反复传代以防止软骨细胞去分化。人耳廓软骨细胞在3D或常规单层条件下培养，分别在第3代和第4代细胞数增加1000倍。在增殖过程中，软骨细胞在三维培养显示出更大的抑制胶原I型（COL 1）和胶原II型（COL 2）的保存比单层。由3D细胞制成的组织工程化软骨也富含COL 2或蛋白多糖，并具有良好的力学性能。3D细胞的优势可能是由于细胞与基质粘附或细胞与细胞接触的微环境与天然软骨的微环境相似。在3D细胞中整合素的上调和钙粘素的下调模仿了天然软骨的表达模式，而不是单层细胞的表达模式。在培养的软骨细胞中，通过小干扰核糖核酸沉默整合素β 1和Ob-cadherin的表达分别导致促进去分化和再分化，表明3D胶原凝胶培养提供了充足的细胞制备并减少了软骨细胞的去分化，这被认为是自体软骨细胞植入的可行策略。
Although autologous chondrocyte implantation has already been in clinical use, chondrocyte dedifferentiation is problematic during proliferation culture. We attempted a three-dimensional (3D) collagen gel culture under chondrocyte proliferation with repeated passaging to prevent the chondrocytes dedifferentiation. Human auricular chondrocytes were cultured in 3D or conventional monolayer conditions, which reached a 1000-fold increase in cell numbers at passages 3 and 4, respectively. During multiplication, the chondrocytes in 3D culture showed greater suppression of collagen type I (COL1) and preservation of collagen type II (COL2) than those in monolayer. Tissue-engineered cartilage made of 3D cells also abundantly accumulated COL2 or proteoglycan and possessed favorable mechanical properties. The advantage of 3D cells may result from the similarity of microenvironments in cell-to-matrix adhesion or cell-to-cell contacts with that of native cartilage. The up-regulation of integrins and down-regulation of cadherins in the 3D cells mimicked the expression pattern of native cartilage, rather than that of monolayer cells. The silencing of integrin beta1 and Ob-cadherin expression by small interfering ribonucleic acid in the cultured chondrocytes led to the promotion of dedifferentiation and redifferentiation, respectively, indicating that the 3D collagen gel culture provided sufficient cell preparation and reduced chondrocyte dedifferentiation, which is regarded as a feasible strategy in autologous chondrocyte implantation.