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骨髓和脂肪组织来源间充质干细胞软骨形成过程中细胞外基质分子的差异表达模式

Differential expression pattern of extracellular matrix molecules during chondrogenesis of mesenchymal stem cells from bone marrow and adipose tissue.

作者信息

Mehlhorn A T, Niemeyer P, Kaiser S, Finkenzeller G, Stark G B, Südkamp N P, Schmal H

机构信息

Department of Orthopaedic and Trauma Surgery, University Medical Center, Albert-Ludwigs University, Freiburg, Germany.

出版信息

Tissue Eng. 2006 Oct;12(10):2853-62. doi: 10.1089/ten.2006.12.2853.

Abstract

Adipose-derived adult stem cells (ADASCs) or bone marrow-derived mesenchymal stem cells (BMSCs) are considered as alternative cell sources for cell-based cartilage repair due to their ability to produce cartilage-specific matrix. This article addresses the differential expression pattern of extracellular matrix (ECM) molecules in BMSCs or ADASCs following chondrogenic differentiation. Human BMSCs or ADASCs were encapsulated in alginate and cultured in TGF-beta1-containing medium for 2 or 3 weeks. Chondrospecific mRNA expression was analyzed and alternative splicing of alpha(1)-procollagen type II mRNA was monitored via reverse transcriptase-polymerase chain reaction (RT-PCR). Corresponding ECM synthesis was demonstrated using immunohistochemistry. After chondroinduction, expression of collagen type II, type X, COMP and aggrecan mRNA was 3-15-fold higher than in ADASCs. The type IIA splicing form of alpha(1)-procollagen type II was expressed in both populations, and the type IIB splicing form was exclusively detected in BMSCs. In response to TGF-beta, collagen type II and X were secreted more strongly by BMSCs than by ADASCs. BMSCs express a more mature phenotype than ADASCs after chondroinduction. TGF-beta1 induces alternative splicing of the alpha(1)-procollagen type II transcript in BMSCs, but not in ADASCs. These findings may direct the development of a cell-specific culture environment either to prevent hypertrophy in BMSCs or to promote chondrogenic maturation in ADASCs.

摘要

脂肪来源的成体干细胞(ADASCs)或骨髓来源的间充质干细胞(BMSCs)因其能够产生软骨特异性基质,被视为基于细胞的软骨修复的替代细胞来源。本文探讨了软骨形成分化后BMSCs或ADASCs中细胞外基质(ECM)分子的差异表达模式。将人BMSCs或ADASCs封装在藻酸盐中,并在含TGF-β1的培养基中培养2或3周。通过逆转录聚合酶链反应(RT-PCR)分析软骨特异性mRNA表达,并监测II型α(1)-前胶原mRNA的可变剪接。使用免疫组织化学证明相应的ECM合成。软骨诱导后,II型、X型胶原、COMP和聚集蛋白聚糖mRNA的表达比ADASCs高3至15倍。II型α(1)-前胶原的IIA型剪接形式在两个群体中均有表达,而IIB型剪接形式仅在BMSCs中检测到。响应TGF-β,BMSCs比ADASCs更强烈地分泌II型和X型胶原。软骨诱导后,BMSCs比ADASCs表现出更成熟的表型。TGF-β1诱导BMSCs中II型α(1)-前胶原转录本的可变剪接,但不诱导ADASCs中的可变剪接。这些发现可能指导细胞特异性培养环境的开发,以防止BMSCs肥大或促进ADASCs的软骨形成成熟。

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