Reproduction Abstracts

Introduction: Poor maternal diet during development induces physiological, metabolic and anatomical alterations in offspring. These may increase immediate survival chances, but can enhance the risk of disease in later life. Maternal low protein diet (LPD) exclusively in pre-implantation development induces changes in endocytosis, postnatal hypertension, cardiovascular dysfunction and alters behaviour in our mouse model. We have previously shown changes in the embryonic stem cells isolated from pre-implantation blastocysts. Here isolate the less studied trophoblast stem cells (TSCs) from embryos, to establish characteristics of these cells as a model of development of the placenta.

Materials and methods: MF1 mice were mated and assigned a LPD or control diet (NPD) for the pre-implantation period. Blastocysts were collected and cultured on feeder cells. After embryo attachment and outgrowth, the outgrowth was disaggregated. TS cell colonies were expanded by passaging every 3–7 days, and frozen after 36 days. The established TS cell lines were studied in karyotyping, determination of the sex by PCR, and gene expression by qPCR and western bloting.

Results and discussion: We have produced a group of TS cell lines from E3.5 embryos of mothers which have received either NPD (18%) or LPD (9%) diets. The karyotyping showed TS cells have a tendency to spontaneous differentiation, with high tendency to show tetraploidy. Unlike ES cells from similarly treated embryos, there is no sex bias in TS lines, however TS cells demonstrate a far greater variability in their culture characteristics, growth rates and stability. This variation does not appear to depend upon the maternal pre-implantation diet.