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血小板裂解液作为心血管组织工程中胎牛血清的自体替代品。

Platelet-lysate as an autologous alternative for fetal bovine serum in cardiovascular tissue engineering.

机构信息

Department of Cardio-Thoracic Surgery, University Medical Center Utrecht , Utrecht, The Netherlands.

出版信息

Tissue Eng Part A. 2010 Apr;16(4):1317-27. doi: 10.1089/ten.TEA.2009.0331.

Abstract

There is an ongoing search for alternative tissue culture sera to engineer autologous tissues, since use of fetal bovine serum (FBS) is limited under Good Tissue Practice guidelines. We compared FBS with human platelet-lysate (PL) in media for in vitro cell culture. A threefold increase in duplication rate was found when human, saphenous vein-derived myofibroblasts were cultured in PL, whereas expression of marker proteins (alpha-smooth muscle actin, vimentin, desmin, and nonmuscle myosin heavy chain) was similar. Heat shock protein 47 mRNA expression was increased in PL cells, and type III collagen fibers were seen on PL-cell monolayers but not on cells cultured in FBS. These results imply a more efficient collagen fiber production. We also found higher levels of proteins involved in tissue repair and collagen remodeling, which could explain increased production of proteases and protease inhibitors by PL cells. Our findings indicate that PL is beneficial due to the increased duplication rate, in addition to the increased matrix production and remodeling. This could lead to production of strong tissue with properly organized collagen fibers, which is important for heart valve tissue engineering.

摘要

目前正在寻找替代组织培养胎牛血清的方法来构建自体组织,因为根据良好组织实践指南,胎牛血清的使用受到限制。我们将胎牛血清与人血小板裂解液(PL)进行了比较,以评估它们在体外细胞培养中的应用。结果发现,人源性大隐静脉来源的肌成纤维细胞在 PL 中培养时,其倍增率增加了三倍,而标志物蛋白(α-平滑肌肌动蛋白、波形蛋白、结蛋白和非肌肉肌球蛋白重链)的表达相似。PL 细胞中的热休克蛋白 47 mRNA 表达增加,PL 细胞单层上可见 III 型胶原纤维,但在 FBS 中培养的细胞上则未见。这些结果表明 PL 能够更有效地产生胶原纤维。我们还发现,PL 细胞中与组织修复和胶原重塑相关的蛋白水平更高,这可以解释 PL 细胞产生更多的蛋白酶和蛋白酶抑制剂。我们的研究结果表明,PL 不仅能够增加细胞的倍增率,还能够增加细胞外基质的产生和重塑,因此对组织工程心脏瓣膜具有重要意义。

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