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Osaka Medical College

3 ARTICLES PUBLISHED IN JoVE

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Cancer Research

One-step Protocol for Evaluation of the Mode of Radiation-induced Clonogenic Cell Death by Fluorescence Microscopy
Daijiro Kobayashi 1, Atsushi Shibata 2, Takahiro Oike 1, Takashi Nakano 1
1Department of Radiation Oncology, Gunma University Graduate School of Medicine, 2Advanced Scientific Research Leaders Development Unit, Gunma University Graduate School of Medicine

Research on ionizing radiation (IR)-induced clonogenic cell death is important for understanding the effects of IR on malignant tumors and normal tissues. Here, we describe a one-step assay for assessing the major modes of IR-induced clonogenic cell death based on morphology of 4',6-diamidino-2-phenylindole dihydrochloride (DAPI)-stained nuclei visualized by fluorescence microscopy.

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Immunology and Infection

Measuring Dengue Virus RNA in the Culture Supernatant of Infected Cells by Real-time Quantitative Polymerase Chain Reaction
Youichi Suzuki 1, Mami Kotoura 1, Shun Yashima 1, Hong Wu 1, Takashi Nakano 1, Kouichi Sano 1
1Department of Microbiology and Infection Control, Osaka Medical College

Real-time quantitative polymerase chain reaction analysis combined with reverse transcription (RT-qPCR) has been widely used to measure the level of RNA virus infections. Here we present a direct RT-qPCR assay, which does not require an RNA purification step, developed for the quantification of several RNA viruses, including dengue virus.

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Bioengineering

Evaluation of Keratinocyte Proliferation on Two- and Three-dimensional Type I Collagen Substrates
Hitomi Fujisaki 1, Sugiko Futaki 2, Kazunori Mizuno 1, Shunji Hattori 1
1Nippi Research Institute of Biomatrix, 2Department of Anatomy and Cell Biology, Osaka Medical College

Here, we present a protocol for the preparation of two different forms of culture substrates utilizing type I collagen. Depending on how collagen is handled, collagen molecules either maintain two-dimensional, non-fibrous form or reassemble into three-dimensional, fibril form. Cell proliferation on type I collagen is drastically affected by fibril formation.

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