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一种基于生物工程的方法,用于开发成纤维细胞生长因子 7 功能化丝胶生物材料,适用于角质细胞的培养。

A Bioengineering Approach for the Development of Fibroblast Growth Factor-7-Functionalized Sericin Biomaterial Applicable for the Cultivation of Keratinocytes.

机构信息

Department of Applied Biology, Kyoto Institute of Technology, Sakyo-ku, Kyoto 606-8585, Japan.

Biomedical Research Center, Kyoto Institute of Technology, Sakyo-ku, Kyoto 606-8585, Japan.

出版信息

Int J Mol Sci. 2022 Sep 1;23(17):9953. doi: 10.3390/ijms23179953.

Abstract

Growth factors, including fibroblast growth factor-7 (FGF-7), are a group of proteins that stimulate various cellular processes and are often used with carriers to prevent the rapid loss of their activities. Sericin with great biocompatibility has been investigated as a proteinaceous carrier to enhance the stability of incorporated proteins. The difficulties in obtaining intact sericin from silkworm cocoons and the handling of growth factors with poor stability necessitate an efficient technique to incorporate the protein into a sericin-based biomaterial. Here, we report the generation of a transgenic silkworm line simultaneously expressing and incorporating FGF-7 into cocoon shells containing almost exclusively sericin. Growth-factor-functionalized sericin cocoon shells requiring simple lyophilization and pulverization processes were successfully used to induce the proliferation and migration of keratinocytes. Moreover, FGF-7 incorporated into sericin-cocoon powder exhibited remarkable stability, with more than 70% of bioactivity being retained after being stored as a suspension at 25 °C for 3 months. Transgenic sericin-cocoon powder was used to continuously supply biologically active FGF-7 to generate a three-dimensionally cultured keratinocyte model in vitro. The outcomes of this study propound a feasible approach to producing cytokine-functionalized sericin materials that are ready to use for cell cultivation.

摘要

生长因子,包括成纤维细胞生长因子 7(FGF-7),是一组刺激各种细胞过程的蛋白质,通常与载体一起使用以防止其活性迅速丧失。具有良好生物相容性的丝胶已被研究作为蛋白质载体,以提高掺入蛋白质的稳定性。从蚕茧中获得完整丝胶以及处理稳定性差的生长因子存在困难,因此需要一种有效的技术将蛋白质掺入丝胶基生物材料中。在这里,我们报告了一种同时表达和将 FGF-7 掺入仅含有丝胶的蚕茧壳中的转基因家蚕系的产生。经过简单的冷冻干燥和粉碎处理的具有生长因子功能的丝胶茧壳成功地用于诱导角质形成细胞的增殖和迁移。此外,掺入丝胶-茧粉中的 FGF-7 表现出显著的稳定性,在 25°C 下以悬浮液形式储存 3 个月后,仍保留超过 70%的生物活性。转基因为丝胶-茧粉持续供应生物活性 FGF-7,以在体外生成三维培养的角质形成细胞模型。这项研究的结果提出了一种可行的方法来生产细胞因子功能化的丝胶材料,这些材料可立即用于细胞培养。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0805/9456417/0349431a1ff5/ijms-23-09953-g001.jpg

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