Kamigyō-ku, Japan
Kamigyō-ku, Japan

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Nishishita N.,Foundation for Biomedical Research and Innovation TRI308 | Nishishita N.,RIKEN | Ijiri H.,Kyoto Institute of Technology | Takenaka C.,Foundation for Biomedical Research and Innovation TRI308 | And 9 more authors.
Biomaterials | Year: 2011

Human leukemia inhibitory factor (LIF) was immobilized into insect virus-derived microcrystals (polyhedra) to generate LIF polyhedra (LIF-PH) that can slowly release LIF into embryonic stem (ES) cell culture media and thus maintain ES cells in an undifferentiated state. Assays of the biological activities of LIF-PH indicated that a single addition of LIF-PH to the ES cell culture medium can support the proliferation of mouse ES and induced pluripotent stem (iPS) cells continuously for 14 days, and suggest that LIF-PH can be successfully used in the place of a periodic addition of recombinant LIF to the media every 2-3 days. The release of LIF protein from LIF-PH was determined by enzyme-linked immunosorbent assay (ELISA). Maintenance of undifferentiated state of mouse ES and iPS cells cultured with LIF-PH was determined by the detection of pluripotency-related biomarkers Oct3/4 and stage-specific embryonic antigen-1 (SSEA-1) through immunostaining and measurement of alkaline phosphatase activity. In this paper, we propose a closed culture system for mass production of ES and iPS cells that utilize a slow-releasing agent of LIF. © 2011 Elsevier Ltd.

Matsushima K.,Foundation for Biomedical Research and Innovation TR1308 | Matsushima K.,Bay Bioscience Corporation | Suyama T.,Foundation for Biomedical Research and Innovation TR1308 | Suyama T.,Bay Bioscience Corporation | And 9 more authors.
Tissue Engineering - Part A | Year: 2010

Expression of the Wnt modulator secreted frizzled related protein 4 (Sfrp4) is upregulated after heart ischemic injury. We show that intramuscular administration of recombinant Sfrp4 to rat heart ischemic injury and recanalization models prevents further deterioration of cardiac function after the ischemic injury. The effect of Sfrp4 persisted for at least 20 weeks when Sfrp4 was administered in a slow release system (Sfrp4-polyhedra) to both acute and subacute ischemic models. The histology of the dissected heart showed that the cardiac wall was thicker and the area of acellular scarring was smaller in Sfrp4-treated hearts than in controls. Increased amounts of both the inactive serine 9-phosphorylated form of glycogen syntase kinase (GSK)-3β and the active form of β-catenin were observed by immunohistology 3 days after lateral anterior descendant ligation in control, but not in Sfrp4-treated hearts. All together, we show that administration of Sfrp4 interferes with canonical Wnt signaling that could mediate the formation of acellular scar and consequently contributes to the prevention of aggravation of cardiac function. © 2010 Mary Ann Liebert, Inc.

Ijiri H.,Kyoto Institute of Technology | Nakatani T.,Kyoto Institute of Technology | Ido H.,Kyoto Institute of Technology | Hamada N.,Kyoto Institute of Technology | And 4 more authors.
Journal of Insect Biotechnology and Sericology | Year: 2010

Certain insect viruses produce stable infectious micro-crystals called polyhedra which function to protect the virus after the death of infected larvae. Polyhedra form within infected cells and contain numerous virus particles embedded in the crystalline lattice of the viral protein polyhedrin. We have previously demonstrated a novel protein engineering technology for the immobilization of protein molecules into Bombyx mori cypovirus (BmCPV) polyhedra. In addition, polyhedra containing foreign proteins have been shown to be highly stable and able to maintain the function of immobilized proteins. In this study, protein kinase C (PKC) was immobilized into polyhedra by using two types of immobilization signals. One immobilization signal was developed by the identification of the BmCPV virus occlusion mechanism into the polyhedra. Another immobilization signal was identified by the atomic structural analysis of the BmCPV polyhedrin. After PKC was fused each with an immobilization signal, the recombinant PKC was immobilized into polyhedra and the enzymatic activities were assayed. These results showed that the activity of PKC immobilized into polyhedra has been maintained and compared with commercially used PKC.

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