Abstract
Conventional reporter gene technology and histological methods cannot routinely be used to track the in vivo behavior of embryonic stem (ES) cells longitudinally after cellular transplantation. Here we describe a protocol for monitoring the in vivo survival, proliferation, and migration of ES cells without necessitating animal sacrifice. Stable ES cell lines containing double fusion (DF; enhanced green fluorescent protein and firefly luciferase) or triple fusion (TF; monomeric red fluorescent protein, firefly luciferase, and herpes simplex virus thymidine kinase) reporter genes can be established within 4-6 weeks by lentiviral transduction followed by fluorescence-activated cell sorting. The cell fate and behavior of these DF or TF ES cells can subsequently be tracked noninvasively by bioluminescence and microPET imaging for a prolonged period of time.