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. 2014 Jan 15;385(2):155-9.
doi: 10.1016/j.ydbio.2013.11.014. Epub 2013 Nov 20.

On the fate of primordial germ cells injected into early mouse embryos

Harry G Leitch  1 , Daiji Okamura  2 , Gabriela Durcova-Hills  3 , Colin L Stewart  4 , Richard L Gardner  5 , Yasuhisa Matsui  2 , Virginia E Papaioannou  6
Affiliations

On the fate of primordial germ cells injected into early mouse embryos

Harry G Leitch et al. Dev Biol. .

Abstract

Primordial germ cells (PGCs) are the founder cells of the germline. Via gametogenesis and fertilisation this lineage generates a new embryo in the next generation. PGCs are also the cell of origin of multilineage teratocarcinomas. In vitro, mouse PGCs can give rise to embryonic germ (EG) cells - pluripotent stem cells that can contribute to primary chimaeras when introduced into pre-implantation embryos. Thus, PGCs can give rise to pluripotent cells in the course of the developmental cycle, during teratocarcinogenesis and by in vitro culture. However, there is no evidence that PGCs can differentiate directly into somatic cell types. Furthermore, it is generally assumed that PGCs do not contribute to chimaeras following injection into the early mouse embryo. However, these data have never been formally published. Here, we present the primary data from the original PGC-injection experiments performed 40 years ago, alongside results from more recent studies in three separate laboratories. These results have informed and influenced current models of the relationship between pluripotency and the germline cycle. Current technologies allow further experiments to confirm and expand upon these findings and allow definitive conclusions as to the developmental potency of PGCs.

Keywords: Germline; Pluripotency; Primordial germ cells.

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Figures

Fig. 1
Fig. 1
Blastocyst injection of PGCs across the decades. (A) Schematic representation of the experimental design for the original PGC blastocyst injection experiments by Papaioannou and Gardner. AP=alkaline phosphatase. (B) Comparison of PGC injection experiments. No evidence of chimaerism was detected at any stage.
Fig. 2
Fig. 2
A modern approach to assessing the pluripotency of PGCs. (A) Phase contrast and fluorescence images showing the isolation of PGCs and somatic cells from E7.5 embryos expressing both mil1-GFP, expressed in PGCs, and a constitutively active CAG-DsRed transgene, expressed in all cells. Scale bar, 100 µm (top row) and 50 µm (bottom two rows). (B) Phase contrast and fluorescence images of blastocysts 2 h after injection with mil1-GFP positive PGCs. Scale bar, 50 µm. (C) Phase contrast and fluorescence images of embryos 24 h after injection of either PGCs or control somatic cells. Mil1-GFP-positive PGCs or GFP-negative somatic cells were injected under the zona pellucida of 8-cell stage embryos. Injected cells can still be visualised by DsRed fluorescence 24 h later. Notably mil1-GFP expression was lost in the injected PGCs. Some fluorescent foci with the appearance of apoptotic fragments are also present. Scale bar, 50 µm.
See this image and copyright information in PMC

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