Highly robust protein production by co-culture of CHO spheroids layered on feeder cells in serum-free medium

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• 作者：Koichi Kutsuzawa (1) (2)
  Toshihiro Suzuki (3) (5)
  Hidehiro Kishimoto (3) (5)
  Akiichi Murakami (4)
  Takachika Azuma (4) (5)
  Ryo Abe (3) (5)
  Hidenori Otsuka (1) (2) (5)
  
• 关键词：Recombinant therapeutic proteins ; Chinese hamster ovary (CHO) cells ; Spheroid ; 3D co ; culture
• 刊名：Colloid & Polymer Science
• 出版年：2014
• 出版时间：April 2014
• 年：2014
• 卷：292
• 期：4
• 页码：839-848
• 全文大小：4,951 KB
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• 作者单位：Koichi Kutsuzawa (1) (2)
  Toshihiro Suzuki (3) (5)
  Hidehiro Kishimoto (3) (5)
  Akiichi Murakami (4)
  Takachika Azuma (4) (5)
  Ryo Abe (3) (5)
  Hidenori Otsuka (1) (2) (5)
  
  1. Department of Applied Chemistry, Faculy of Science, Tokyo University of Science, 1-3 Kagurazaka, Shinjuku-ku, Tokyo, 162-8601, Japan
  2. Center for Colloid and Interface Science, Research Institute for Science and Technology, Tokyo University of Science, 1-3 Kagurazaka, Shinjuku-ku, Tokyo, 162-8601, Japan
  3. Division of Immunobiology, Research Institute for Biomedical Sciences, Tokyo University of Science, Chiba, 278-0022, Japan
  5. Center for Technologies against Cancer, Research Institute for Science and Technology, Tokyo University of Science, Chiba, 278-0022, Japan
  4. Division of Biosignaling, Research Institute for Biomedical Sciences, Tokyo University of Science, Chiba, 278-0022, Japan
  
• ISSN：1435-1536

文摘

Recombinant Chinese hamster ovary (rCHO) cells have been the most commonly used mammalian host for large-scale commercial production of therapeutic proteins. Although recent advances in 3D culture of rCHO cells is preferred to 2D monolayer culture for highly productive and robust expression of therapeutic proteins, there exists still limitation for efficient protein production. Therefore, a new cell culture system is essentially required for an efficient protein production. Here, we report on a new 3D cell culture system as a spheroid cell culture on the micropattern array for efficient production of protein by CHO cells. Particularly, cocultivation of CHO spheroids with bovine aortic endothelial cells (BAEC) as a feeder layer cells was essential to stably increase a protein production. We investigated the co-culture mechanism of functional enhancement with respect to the cell–cell interactions. Functional comparison between 2D and 3D co-cultures suggested the preferred configuration as spheroid for higher protein production. Specifically, to estimate the effect of respective cell constitution in co-cultured spheroids on the protein production per CHO cell, the number of viable cells in cell proliferation was determined with culture periods. These studies demonstrated the significant role of micropatterned BAEC as a feeder layer for the retained formation of CHO spheroids, resulting in predominantly enhanced production of proteins, although the functional enhancement of CHO cells was obtained by co-culture with BAECs in both 2D and 3D configurations. Thus, heterotypic cell communications that play indispensable roles in increasing CHO functions should be properly obtained in 3D cell configurations. Significantly, these spheroids in the serum-free medium drastically enhanced protein expression level up to sevenfold compared with CHO monospheroids, suggesting that a suitable culture conditions for heterotypic cell–cell interactions would allow improved protein secretion to occur unimpeded.