Osaka Medical College

3 ARTICLES PUBLISHED IN JoVE

Cancer Research

One-step Protocol for Evaluation of the Mode of Radiation-induced Clonogenic Cell Death by Fluorescence Microscopy

Daijiro Kobayashi¹, Atsushi Shibata², Takahiro Oike¹, Takashi Nakano¹

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.

Immunology and Infection

Measuring Dengue Virus RNA in the Culture Supernatant of Infected Cells by Real-time Quantitative Polymerase Chain Reaction

Youichi Suzuki¹, Mami Kotoura¹, Shun Yashima¹, Hong Wu¹, Takashi Nakano¹, Kouichi Sano¹

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.

Bioengineering

Evaluation of Keratinocyte Proliferation on Two- and Three-dimensional Type I Collagen Substrates

Hitomi Fujisaki¹, Sugiko Futaki², Kazunori Mizuno¹, Shunji Hattori¹

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.