Accedi

Manipal Academy of Higher Education

3 ARTICLES PUBLISHED IN JoVE

Bioengineering

A Simple Microfluidic Chip for Long-Term Growth and Imaging of Caenorhabditis elegans

Jyoti Dubey^1,2,3,4, Sudip Mondal^1,5, Sandhya P. Koushika²

¹National Centre for Biological Sciences, TIFR, ²Department of Biological Sciences, TIFR, ³InSTEM, ⁴Manipal Academy of Higher Education, ⁵Department of Mechanical Engineering, The University of Texas at Austin

The protocol describes a simple microfluidic chip design and microfabrication methodology used to grow C. elegans in presence of a continuous food supply for up to 36 h. The growth and imaging device also enables intermittent long-term high-resolution imaging of cellular and sub-cellular processes during development for several days.

Biology

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. Karuppusamy^1,2, Prathibha Babu^1,2, Manoj Kumar K. Alagiri^1,2, 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.

Developmental Biology

Generation of Retinal Organoids from Healthy and Retinal Disease-Specific Human-Induced Pluripotent Stem Cells

Sudipta Mahato^*1,2, Trupti Agrawal^*1,2, Divya Pidishetty^*1,2, Savitri Maddileti¹, Vinay Kumar Pulimamidi^1,3, Indumathi Mariappan¹

¹Centre for Ocular Regeneration, Prof. Brien Holden Eye Research Centre, LV Prasad Eye Institute, ²Manipal Academy of Higher Education, ³Schepens Eye Research Institute, Massachusetts Eye and Ear, Harvard Medical School

This protocol describes an efficient method of differentiating hiPSCs into eye field clusters and generating neuro-retinal organoids using simplified culture conditions involving both adherent and suspension culture systems. Other ocular cell types, such as the RPE and corneal epithelium, can also be isolated from mature eye fields in retinal cultures.