Pohang University of Science and Technology

6 ARTICLES PUBLISHED IN JoVE

Bioengineering

Fully Automated Centrifugal Microfluidic Device for Ultrasensitive Protein Detection from Whole Blood

Yang-Seok Park¹, Vijaya Sunkara¹, Yubin Kim¹, Won Seok Lee^1,2, Ja-Ryoung Han^1,3, Yoon-Kyoung Cho^1,4

¹Department of Biomedical Engineering, School of Life Sciences, Ulsan National Institute of Science and Technology (UNIST), UNIST-gil 50, Ulsan, Republic of Korea, ²Agency for Defense Development (ADD), Daejeon, Republic of Korea, ³KOGAS (Korea Gas Corporation) Research Institute, ⁴Center for Soft and Living Matter, Institute for Basic Science (IBS)

This protocol demonstrates how to achieve femto molar detection sensitivity of proteins in 10 µL of whole blood within 30 min. This can be achieved by using electrospun nanofibrous mats integrated in a lab-on-a-disc, which offers high surface area as well as effective mixing and washing for enhanced signal-to-noise ratio.

Biology

Observation and Quantification of Telomere and Repetitive Sequences Using Fluorescence In Situ Hybridization (FISH) with PNA Probes in Caenorhabditis elegans

Beomseok Seo¹, Junho Lee^1,2,3

¹Institute of Molecular Biology and Genetics (IMBG), Seoul National University, ²Department of Biological Sciences, Seoul National University, ³Department of Biophysics and Chemical Biology, Seoul National University

We report a concise procedure of fluorescence in situ hybridization (FISH) in the gonad and embryos of Caenorhabditis elegans for observing and quantifying repetitive sequences. We successfully observed and quantified two different repetitive sequences, telomere repeats and template of alternative lengthening of telomeres (TALT).

Biochemistry

Studying Protein Import into Chloroplasts Using Protoplasts

Junho Lee¹, Hyangju Kang¹, Inhwan Hwang^1,2

¹Division of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology, ²Department of Life Sciences, Pohang University of Science and Technology

Here we describe a protocol to express proteins into protoplasts by using PEG-mediated transformation method. The method provides easy expression of proteins of interest, and efficient investigation of protein localization and the import process for various experimental conditions in vivo.

Bioengineering

Pancreatic Tissue-Derived Extracellular Matrix Bioink for Printing 3D Cell-Laden Pancreatic Tissue Constructs

Jaewook Kim^*1, Myungji Kim^*2, Dong Gyu Hwang², In Kyong Shim³, Song Cheol Kim^3,4, Jinah Jang^1,2,5

¹Department of Mechanical Engineering, Pohang University of Science and Technology, ²School of Interdisciplinary Bioscience and Bioengineering, Pohang University of Science and Technology, ³Asan Institute for Life Science, University of Ulsan College of Medicine and Asan Medical Center, ⁴Division of Hepato-Biliary and Pancreatic Surgery, Department of Surgery, University of Ulsan College of Medicine and Asan Medical Center, ⁵Department of Creative IT Engineering, Pohang University of Science and Technology

Decellularized extracellular matrix (dECM) can provide suitable microenvironmental cues to recapitulate the inherent functions of target tissues in an engineered construct. This article elucidates the protocols for the decellularization of pancreatic tissue, evaluation of pancreatic tissue-derived dECM bioink, and generation of 3D pancreatic tissue constructs using a bioprinting technique.

Cancer Research

Culture, Manipulation, and Orthotopic Transplantation of Mouse Bladder Tumor Organoids

Yubin Kim^*1, Juhee Lee^*1, SungEun Kim^*1, Kunyoo Shin¹

¹Department of Life Sciences, Pohang University of Science and Technology

This protocol provides detailed experimental steps to establish a three-dimensional in vitro culture of bladder tumor organoids derived from carcinogen-induced murine bladder cancer. Culture methods including passaging, genetic engineering, and orthotopic transplantation of tumor organoids are described.

Bioengineering

3D Cell-Printed Hypoxic Cancer-on-a-Chip for Recapitulating Pathologic Progression of Solid Cancer

Wonbin Park^*1, Mihyeon Bae^*1, Minseon Hwang¹, Jinah Jang², Dong-Woo Cho¹, Hee-Gyeong Yi^1,3

¹Department of Mechanical Engineering, Pohang University of Science and Technology (POSTECH), ²Department of Creative IT Engineering, Pohang University of Science and Technology (POSTECH), ³Department of Rural and Biosystems Engineering, College of Agriculture and Life Sciences, Chonnam National University

Hypoxia is a hallmark of tumor microenvironment and plays a crucial role in cancer progression. This article describes the fabrication process of a hypoxic cancer-on-a-chip based on 3D cell-printing technology to recapitulate a hypoxia-related pathology of cancer.