Journal: Journal of visualized experiments : JoVE

Article Title: Establishing a High Throughput Epidermal Spheroid Culture System to Model Keratinocyte Stem Cell Plasticity

doi: 10.3791/62182

Figure Lengend Snippet: Outlines PCR primer sequences used for the detection of select genes involved in neonatal keratinocyte plasticity.

Article Snippet: Use GAPDH as an internal control ( ). table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Name of Material/ Equipment Company Catalog Number Comments/Description MCDB 153-LB basal media Sigma-Aldrich M7403 MCDB 153-LB basal media w/ HEPES buffer KSFM-scm ThermoFisher Scientific 17005042 Supplemented with 1% Penicillin/Streptomycin, 20 ng/ml EGF, 10 ng/ml basic fibroblast growth factor, 0.4% bovine serum albumin (BSA), and 4 μg/ml insulin Human Basic Fibroblast Growth Factor (hFGF basic/FGF2) Cell Signaling Technology 8910 Human Epidermal Growth Factor (hEGF) Cell Signaling Technology 8916 Human Insulin Millipore Sigma 9011-M Thermo Scientific™ Sterile Single Use Vacuum Filter Units Thermo Scientific 09-740-63D FITC-conjugated anti-integrinα6 Abcam ab30496 PE-conjugated anti-EGFR ( San Jose, CA; catalog number ) BD Pharmingen 555997 Dispase Sigma-Aldrich D4818 NEST Scientific 1-Well Cell Culture Chamber Slide, BLACK Walls on Glass Slide, 6/PK, 12/CS Stellar Scientific NST230111 P63 Thermo Scientific 703809 1:200 dilution Cytokeratin 14 Santa Cruz Biotechnology sc-53253 1:200 dilution All Prep DNA/RNA Mini Kit Qiagen 80204 iScript cDNA Synthesis Kit Bio-Rad 1708890 iQ SYBR Green Supermix (Bio-Rad) Bio-Rad 1708880 Promega TransFast kit Promega E2431 pMSCV-IRES-EGFP plasmid vector Addgene 20672 Open in a separate window Outlines PCR primer sequences used for the detection of select genes involved in neonatal keratinocyte plasticity.

Techniques: Cell Culture, SYBR Green Assay, Plasmid Preparation