Muraya Koji, Kawasaki Tomoyuki, Yamamoto Takeshi, Akutsu Hidenori
Bioscience and Technology Development Center, FUJIFILM Corporation, Kanagawa, Japan.
Department of Reproductive Medicine, National Center for Child Health and Development, Tokyo, Japan.
Biores Open Access. 2019 Nov 22;8(1):210-218. doi: 10.1089/biores.2019.0012. eCollection 2019.
Mesenchymal stromal cells (MSCs) have considerable potential for a wide range of clinical applications and regenerative medicine and cell therapy. As a consequence, there is considerable interest in developing robust culture methods for producing large number of MSCs for use in repair of injured tissues or treatment of diseases. In general, tissue culture plates or flasks that have been precoated with substrates derived from animal tissues are used in the production of MSCs. However, these substrates can potentially cause serious problems due to contamination of the MSCs with animal-derived components. In this study, we evaluated the use of a type I collagen-based recombinant peptide (RCP) for MSC culture in an attempt to avoid the problems associated with animal cell-derived substances. This RCP is xeno free, has an increased RGD (Arg-Gly-Asp) sequence, and has high molecular weight uniformity. The effect of RCP on promotion of cellular adhesion and proliferation of MSCs was investigated in cultures in which RCP was included in the culture medium. The effects of RCP on promotion of cellular adhesion and proliferation of MSCs were investigated by comparing cultures in which the additive was present in the culture medium and those where the culture plates were coated with RCP. In addition, changes in gene expression profiles during cell culture were monitored by real time-polymerase chain reaction. Our analyses showed that RCP enhanced cellular adhesion and proliferation in cultures in which the additive was included in the culture medium. Our findings indicate that adding RCP to the culture medium could save time and cost in MSC culture. Our gene expression analysis indicated that RCP enhanced expression of genes encoding proteins associated with the extracellular matrix and cell adhesion.
间充质基质细胞（MSCs）在广泛的临床应用、再生医学以及细胞治疗方面具有巨大潜力。因此，人们对开发强大的培养方法以大量生产MSCs用于受损组织修复或疾病治疗有着浓厚兴趣。一般来说，在MSCs的生产过程中使用的是预先涂覆有源自动物组织的底物的组织培养板或培养瓶。然而，这些底物可能会因MSCs被动物源性成分污染而引发严重问题。在本研究中，我们评估了一种基于I型胶原蛋白的重组肽（RCP）用于MSCs培养的情况，试图避免与动物细胞源性物质相关的问题。这种RCP不含异种成分，具有增加的RGD（精氨酸 - 甘氨酸 - 天冬氨酸）序列，并且具有高分子量均匀性。在培养基中加入RCP的培养物中，研究了RCP对促进MSCs细胞黏附和增殖的作用。通过比较培养基中存在添加剂的培养物和培养板涂有RCP的培养物，研究了RCP对促进MSCs细胞黏附和增殖的作用。此外，通过实时聚合酶链反应监测细胞培养过程中基因表达谱的变化。我们的分析表明，在培养基中加入添加剂的培养物中，RCP增强了细胞黏附和增殖。我们的研究结果表明，在MSCs培养中向培养基中添加RCP可以节省时间和成本。我们的基因表达分析表明，RCP增强了与细胞外基质和细胞黏附相关的蛋白质编码基因的表达。
