Highly efficient gene transfer into primate embryonic stem cells with a simian lentivirus vector

Area of Science:

• Stem cell biology
• Gene therapy
• Primate research

Background:

• Primate embryonic stem (ES) cells hold significant potential for research and therapeutics.
• Stable genetic modification of these cells is essential for their broader application.
• Previous work established cynomolgus monkey ES cell lines.

Purpose of the Study:

• To investigate the efficiency and stability of lentiviral gene transfer into cynomolgus monkey ES cells.
• To compare gene transfer efficiency between primate and mouse ES cells using simian immunodeficiency virus (SIV)-based vectors.
• To assess the utility of green fluorescent protein (GFP) as a reporter for monitoring gene expression in transduced ES cells.

Main Methods:

• Transduction of cynomolgus ES cells with an SIV-based lentivirus vector encoding the GFP gene.
• Monitoring of GFP expression levels and stability over time (5 months) without selection.
• Observation of GFP expression during embryoid body formation.
• Comparative transduction of mouse ES cells with the same SIV-based vector.

Main Results:

• Approximately 90% of cynomolgus ES cells exhibited stable GFP expression for 5 months post-transduction.
• High GFP expression was maintained during embryoid body formation, indicating successful differentiation.
• SIV-based lentivirus vectors demonstrated higher transduction efficiency in primate ES cells compared to mouse ES cells.
• GFP served as an effective reporter for tracking transduced cells and their differentiation.

Conclusions:

• Lentiviral gene transfer is a highly efficient and stable method for genetic modification of primate ES cells.
• SIV-based vectors show preferential transduction of primate ES cells over mouse ES cells.
• This technique facilitates monitoring of ES cell behavior and holds promise for future research and therapeutic applications in non-human primates.