3 articles published in JoVE
CRISPR/Cas9 Gene Editing of Hematopoietic Stem and Progenitor Cells for Gene Therapy Applications Vigneshwaran Venkatesan*1,2, Abisha Crystal Christopher*1,3, Karthik V. Karuppusamy1,2, Prathibha Babu1,2, Manoj Kumar K. Alagiri1,2, Saravanabhavan Thangavel1 1Centre for Stem Cell Research (CSCR), A unit of InStem Bengaluru, Christian Medical College campus, 2Manipal Academy of Higher Education, 3Thiruvalluvar University The present protocol describes an optimized hematopoietic stem and progenitor cell (HSPC) culture procedure for the robust engraftment of gene-edited cells in vivo.
Pooled shRNA Library Screening to Identify Factors that Modulate a Drug Resistance Phenotype Saswati Das1,2, Raveen Stephen Stallon Illangeswaran1,2, Smitha Ijee2,3, Sanjay Kumar2,3, Shaji Ramachandran Velayudhan1,2,3, Poonkuzhali Balasubramanian1,2,3 1Department of Clinical Haematology, Christian Medical College, 2Department of Biotechnology, Thiruvalluvar University, 3Centre for Stem Cell Research, Christian Medical College High-throughput RNA interference (RNAi) screening using a pool of lentiviral shRNAs can be a tool to detect therapeutically relevant synthetic lethal targets in malignancies. We provide a pooled shRNA screening approach to investigate the epigenetic effectors in acute myeloid leukemia (AML).
Human Pluripotent Stem Cell Culture on Polyvinyl Alcohol-Co-Itaconic Acid Hydrogels with Varying Stiffness Under Xeno-Free Conditions Tzu-Cheng Sung*1, Hsing-Fen Li*1, Akon Higuchi1,2, Qing-Dong Ling3,4, Jia-Sin Yang1, Yeh-Chia Tseng1, Chih-Hsien Pan Pan1, Abdullah A. Alarfaj2, Murugan A. Munusamy2, Suresh Kumar5, Shih-Tien Hsu6, Kadarkarai Murugan7,8 1Department of Chemical and Materials Engineering, National Central University, 2Department of Botany and Microbiology, King Saud University, 3Cathay Medical Research Institute, Cathay General Hospital, 4Graduate Institute of Systems Biology and Bioinformatics, National Central University, 5Department of Medical Microbiology and Parasitology, Universiti Putra Malaysia, 6Department of Internal Medicine, Taiwan Landseed Hospital, 7Department of Zoology, Bharathiar University, 8Thiruvalluvar University A protocol is presented to prepare polyvinyl alcohol-co-itaconic acid hydrogels with varying stiffness, which were grafted with and without oligopeptides, to investigate the effect of the stiffness of biomaterials on the differentiation and proliferation of stem cells. The stiffness of the hydrogels was controlled by the crosslinking time.