Isolation of mouse primordial germ cells

Abstract

Primordial germ cells (PGCs) were obtained from fetal mouse gonads of both sexes 12 1 2 and 13 1 2 days post coitum (dpc), either by collagenase treatment, or by mechanical procedures with or without prior EDTA treatment. With mechanical procedures alone, yield was relatively low and many of the cells released were dead. After EDTA treatment, both yield and viability were significantly improved. Collagenase treatment gave the best yield of cells, since the entire gonad was disaggregated, but contamination with somatic cells was substantial, and the adhesive properties of the germ cells were altered by the treatment. When cells released following EDTA treatment were fractionated on a simple Percoll gradient, several thousand viable PGCs per 13 1 2 day gonad could be obtained in 2–3 h, with not more than 10–20% somatic cell contamination.