Source: Hanna, C., et al. Labeling of Breast Cancer Patient-derived Xenografts with Traceable Reporters for Tumor Growth and Metastasis Studies. J. Vis. Exp. (2016).
The following video describes a technique of transduction to label (patient-derived xenografts) PDX tumor cells, which is used for introducing lentivirus expressing green-fluorescent protein and luciferase reporters into the tumor cell in vitro.
1. Transduction of PDX-derived Tumor Cells
2. Evaluation of Transduction Efficiency and Re-implantation of Labeled Cells in Immunocompromised Mice
Figure 1: Tracking Transduction Efficiency in Dissociated PDX Cells. A) PDX-dissociated cells enriched for human epithelial cells (Lin+ depletion) 24 hr after transduction with pSIH1-H1-copGFP-T2A-puro lentiviral particles. B) PDX-dissociated cells from a separate experiment, without epithelial cell enrichment, 72 hr after transduction with pHAGE-EF1aL-luciferase-UBC-GFP-W. Both panels show live cells. BF: Bright field images, bar represents 50 µm
The authors have nothing to disclose.
DMEM/F12 (1:1) | Hyclone | SH30023.01 | |
bFGF | BD Biosciences | 354060 | |
EGF | BD Biosciences | 354001 | |
Heparin | Sigma | H4784 | |
B27 | Gibco/Thermo Fisher | 17504-44 | |
Anti-fungi-antibiotics | Hyclone | SV30010 | |
HBSS Red Ca2+/Mg2+ free | Hyclone | SH30031.02 | |
Hepes | |||
Cultrex | Cultrex | 3433-005-01 | Basement Matrix Extract (BME) |
30 °C shaker | NewBrunswick Scientific CO. INC | Series 25 Incubator Shaker |