March 2016 - This Month in JoVE: RNA interference in Mosquitoes, iPSCs Derived from Nasal Epithelia, Air Sampling of Atmospheric Aerosols, and Autologous Micro-grafts for Skin Lesions

Protocol

Tissue Characterization after a New Disaggregation Method for Skin Micro-Grafts Generation

Valeria Purpura¹, Elena Bondioli¹, Antonio Graziano², Letizia Trovato², Davide Melandri¹, Martina Ghetti¹, Andrea Marchesini³, Maria Gabriella Cusella De Angelis^4,5, Laura Benedetti^4,5, Gabriele Ceccarelli^4,5, Michele Riccio³

¹Burns Centre and Emilia Romagna Regional Skin Bank, ²Human Brain Wave srl, ³Plastic and Reconstructive Surgery, AOU “Ospedali Riuniti”, ⁴Department of Public Health, Experimental Medicine, Anatomy and Forensic, University of Pavia, ⁵C.H.T Centre for Health Technologies, University of Pavia

The protocol describes a new method to disaggregate human tissues and to create autologous micro-grafts that, combined with collagen sponges, give rise to human bio-complexes ready to use in the treatment of skin lesions. Further, this system preserves cell viability of micro-grafts at different times after mechanical disaggregation.

Air-sampled Filter Analysis for Endotoxins and DNA Content

Naama Lang-Yona^1,2, Yinon Mazar¹, Michal Pardo¹, Yinon Rudich¹

¹Department of Earth and Planetary Sciences, Weizmann Institute of Science, ²Multiphase Chemistry Department, Max Planck Institute

Two complementary analyses of atmospheric biological particles from air sampled filters are described herein: the extraction and detection of endotoxin, and of DNA.

RNAi Trigger Delivery into Anopheles gambiae Pupae

Kimberly Regna¹, Rachel M. Harrison¹, Shannon A. Heyse¹, Thomas C. Chiles¹, Kristin Michel², Marc A. T. Muskavitch^1,3

¹Biology Department, Boston College, ²Division of Biology, Kansas State University, ³Discovery Research, Biogen

RNA interference (RNAi) is an extremely valuable tool for uncovering gene function. However, the ability to target genes using RNAi during pre-adult stages is limited in the major human malaria vector Anopheles gambiae. We describe an RNAi protocol to reduce gene function via direct injection during pupal development.

Cultivate Primary Nasal Epithelial Cells from Children and Reprogram into Induced Pluripotent Stem Cells

Ashley Ulm¹, Christopher N. Mayhew², Jason Debley³, Gurjit K. Khurana Hershey⁴, Hong Ji^1,4

¹Pyrosequencing Core, Cincinnati Children's Hospital, ²Division of Developmental Biology, Cincinnati Children's Hospital, ³Division of Pulmonary Medicine, Seattle Children's Hospital, ⁴Division of Asthma Research, Cincinnati Children's Hospital

This publication demonstrates methods for successful sampling and culture of nasal epithelial mucosa from children, and reprogramming these cells to induced Pluripotent Stem Cells (iPSCs).