Molecular and cellular characterization during chondrogenic differentiation of adipose tissue-derived stromal cells in vitro and cartilage formation in vivo
Molecular and cellular characterization during chondrogenic differentiation of adipose tissue-derived stromal cells in vitro and cartilage formation in vivo
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DOI：
10.1038/cr.2008.257
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发表时间：
2008-08
期刊：
影响因子：
44.1
通讯作者：
Yunfeng Lin;E. Luo;Xizhe Chen;Lei Liu;Zheng-bin Yan;Zhi-yong Li;Wei Tang;Xiao-hui Zheng;W. Tian
中科院分区：
文献类型：
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作者：
Yunfeng Lin;E. Luo;Xizhe Chen;Lei Liu;Zheng-bin Yan;Zhi-yong Li;Wei Tang;Xiao-hui Zheng;W. Tian
人脂肪组织是间充质干细胞（MSCs）的有效来源，在肌肉骨骼组织工程研究中具有广泛的分化潜力。干细胞群，称为加工抽脂（PLA）细胞，可以从人抽脂中分离出来，并且很容易在体外扩增。本研究旨在确定PLA细胞在体外成软骨分化和体内软骨形成过程中的分子和细胞特征。在体外高密度单层成软骨培养基中培养，可诱导成软骨谱系。为了确定它们在体内形成软骨的能力，我们将海藻酸盐凝胶中的诱导细胞皮下植入裸鼠长达20周。对诱导细胞和不同时间间隔的裸鼠标本进行组织学和免疫组化分析，显示明显的软骨表型，特异性细胞外基质（ECM）染色阳性。相关的，RT-PCR和western blot结果证实了软骨分化过程中的特征分子，即II型胶原、SOX9、软骨寡聚蛋白（COMP）和软骨特异性蛋白多糖聚集蛋白的表达。同时，在体外诱导和体内软骨组织形成过程中，X型胶原合成水平较低，I型胶原生成减少。这些被诱导形成工程软骨的细胞在体内能保持稳定的表型，在20周内没有出现肥大的迹象，但当它们作为单层培养时，在诱导后约10周的后期出现了增生性前改变。因此，人类脂肪组织可能是一种新的、丰富的多能干细胞来源，能够进行软骨形成和形成工程软骨。
Human adipose tissue is a viable source of mesenchymal stem cells (MSCs) with wide differentiation potential for musculoskeletal tissue engineering research. The stem cell population, termed processed lipoaspirate (PLA) cells, can be isolated from human lipoaspirates and expanded in vitro easily. This study was to determine molecular and cellular characterization of PLA cells during chondrogenic differentiation in vitro and cartilage formation in vivo. When cultured in vitro with chondrogenic medium as monolayers in high density, they could be induced toward the chondrogenic lineages. To determine their ability of cartilage formation in vivo, the induced cells in alginate gel were implanted in nude mice subcutaneously for up to 20 weeks. Histological and immunohistochemical analysis of the induced cells and retrieved specimens from nude mice at various intervals showed obviously cartilaginous phenotype with positive staining of specific extracellular matrix (ECM). Correlatively, results of RT-PCR and western blot confirmed the expression of characteristic molecules during chondrogenic differentiation namely collagen type II, SOX9, cartilage oligomeric protein (COMP) and the cartilage-specific proteoglycan aggrecan. Meanwhile, there was low level synthesis of collagen type X and decreasing production of collagen type I during induction in vitro and formation of cartilaginous tissue in vivo. These cells induced to form engineered cartilage can maintain the stable phenotype and indicate no sign of hypertrophy in 20 weeks in vivo, however, when they cultured as monolayers, they showed prehypertrophic alteration in late stage about 10 weeks after induction. Therefore, it is suggested that human adipose tissue may represent a novel plentiful source of multipotential stem cells capable of undergoing chondrogenesis and forming engineered cartilage.