Journal: PLoS ONE

Article Title: Decellularized Wharton’s Jelly from human umbilical cord as a novel 3D scaffolding material for tissue engineering applications

doi: 10.1371/journal.pone.0172098

Figure Lengend Snippet: A) Mice with cranial defect, B) mice with cranial defect and DWJM, C) mice with cranial defect and DWJM 14 days post-surgery. Arrows in A represent the defect, B shows the DWJM and C is the defect and DWJM 14 days post-surgery. D) IVIS imaging of the mice post—surgery—1) Mice with DWJM 24 hours post- surgery; 2–6) designates mice 14 days after the surgeries. D2 is mice without any intervention, D3 and D4 are mice with the defect alone, and D5—D6 represent mice with defect and DWJM. The red circles indicate the defect sites and the inset images are a higher magnification of the defect site in mice. The green fluorescence signal at the defect site signifies the migration of the GFP positive cells into the defect . Images E-J represent the histology images of bone specimen with DWJM 14 days post-surgery, with image E) hematoxylin-eosin stained (H&E) section of DWJM tissue specimen 24 hours post-surgery, and image F depicts GFP immunohistochemistry staining of the same. Images G-J represent DWJM sample 14 days post-surgery with G, H and I being H&E stained sections of DWJM viewed at different magnifications as indicated in the figure. J represents the GFP immunohistochemistry of the section in image I. The arrows in image F, J represent GFP positive cells.

Article Snippet: Green fluorescent protein (GFP) immunohistochemistry staining was performed by incubation with primary monoclonal goat-anti GFP antibody (1:100 dilution for 45 minutes), followed by secondary MACH2 rabbit antibody (CST, Cell Signaling Technology, Danvers, MA) MACH 2 for 45 minutes on a Clinical intelliPATH FLX automated slide stainer.

Techniques: Imaging, Fluorescence, Migration, Staining, Immunohistochemistry