Pluripotent Stem Cells In Vitro from Human Primordial Germ Cells

In the mouse, embryonic germ (EG) cell lines can be generated from primordial germ cells (PGCs) �recovered from the genital ridge of the developing fetus. These EG cells have the capacity of self-renewal, are pluripotent, and have the potential to differentiate to other cell types of the body similar to embryonic stem cells (ESCs). In this chapter, we describe a simple procedure to derive and keep in culture putative human EG (hEG) cells. Fetal material (5–8 weeks’ gestation) was obtained following full local ethical approval and patient consent. Fetal gonads were recovered by dissection and treated with collagenase type IV. Cells were washed by centrifugation and cocultured with inactivated mouse embryonic fibroblasts (MEFs) feeder layer in different conditions. These cell lines can be characterized with specific markers. It was not possible to maintain an undifferentiated hEG cell line for more than 30 cell passages.