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首页> 外文期刊>Tissue engineering, Part A >Human adipose-derived stem cells contribute to chondrogenesis in coculture with human articular chondrocytes
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Human adipose-derived stem cells contribute to chondrogenesis in coculture with human articular chondrocytes

机译:人脂肪干细胞与人关节软骨细胞共培养有助于软骨生成

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Adipose tissue is easily available and contains high numbers of stem cells that are capable for chondrogenic differentiation. We hypothesize that a partial substitution of chondrocytes with autologous adipose-derived stem cells (ASC) might be a possible strategy to reduce the number of chondrocytes needed in matrix-associated autologous chondrocyte transplantation. To lay the ground, in vitro coculture experiments were performed using human chondrocytes and human ASC. Chondrocytes were obtained from donors undergoing matrix-associated autologous chondrocyte transplantation. ASC were isolated from liposuction material. Chondrocytes and ASC were seeded either in fibrin (Tisseel?; Baxter, Vienna, Austria) or collagen matrix (Tissue Fleece?; Baxter, Unterschleissheim, Germany). RNA for quantitative reverse transcriptase (RT)-polymerase chain reaction was isolated after 2 weeks of culture in chondrogenic medium, and after 4 weeks samples were processed for histology. Related to the number of chondrocytes used, coculture with ASC led to strong increase in collagen type IX mRNA expression, which is an indicator for long-term stability of cartilage. Moderate upregulation was shown for SOX9, aggrecan, melanoma inhibitory activity, cartilage link protein 1, and cartilage oligomeric matrix protein mRNA. However, expression of collagen I and collagen II indicates the synthesis of fibrous tissue, which might be due to the use of dedifferentiated chondrocytes. Tisseel provided slightly better chondrogenic conditions than Tissue Fleece. These data support the possibility to take advantage of ASC in cartilage regeneration in conjunction with autologous chondrocytes.
机译:脂肪组织容易获得,并且含有大量能够分化为软骨的干细胞。我们假设用自体脂肪干细胞(ASC)部分替代软骨细胞可能是减少基质相关自体软骨细胞移植所需软骨细胞数量的可能策略。为了奠定基础,使用人软骨细胞和人ASC进行了体外共培养实验。软骨细胞获自接受基质相关自体软骨细胞移植的供体。从抽脂材料中分离出ASC。将软骨细胞和ASC接种在纤维蛋白(Tisseel?; Baxter,维也纳,奥地利)或胶原蛋白基质(Tissue Fleece?; Baxter,Unterschleissheim,德国)中。在软骨形成培养基中培养2周后,分离出用于定量逆转录酶(RT)-聚合酶链反应的RNA,并在4周后对样品进行组织学处理。与使用的软骨细胞数量相关,与ASC共培养导致IX型胶原mRNA表达的强烈增加,这是软骨长期稳定性的指标。对SOX9,聚集蛋白聚糖,黑色素瘤抑制活性,软骨连接蛋白1和软骨寡聚基质蛋白mRNA表达适度上调。然而,胶原蛋白I和胶原蛋白II的表达表明纤维组织的合成,这可能是由于使用去分化的软骨细胞所致。 Tisseel提供的软骨生成条件比组织绒毛略好。这些数据支持在自体软骨细胞联合软骨再生中利用ASC的可能性。

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