Introduction: The developmental origins of health and disease (DOHaD) concept proposes that maternal environment during pregnancy may influence offspring health and predispose to chronic disease risk in later life. Our previous mouse in vivo studies showed that maternal low protein diet (LPD) and advanced maternal age (AMA) programme the preimplantation embryo to adult disease, notably cardiometabolic dysfunction. To test this hypothesis and understand the mechanistic basis of periconceptional programming, we derive mESCs from LPD and AMA models.
Methods: MESCs were derived from blastocysts collected from mothers fed LPD (9% casein) and normal protein diet (NPD, 18% casein) only during E3.5 and AMA model of old (78 months) and young (78 weeks) dams.
Results and discussion: LPD lines showed reduced derivation efficiency (20%, P<0.0155 blastocyst/ mother), Nanog: Oct4 expressing cells in outgrowths (P<0.05) and increased apoptotic cells (P<0.05) than NPDs. Cell signaling activity was similar for Akt, Stat3 and p38 pathways but ERK 1/2 was reduced in activity in LPD lines (P<0.05) suggesting reduced MAPK survival signaling may contribute to LPD ESC enhanced apoptosis. Global metabolomics profile of LPD lines depicted altered glucose metabolism, fatty acid homeostasis and ascorbate utilization. These variations indicate stress management strategies of mESCs to combat detrimental effects of LPD induced during (and preserved since) early embryo programming (E3.5). AMA derived lines showed similar embryo and ESC derivation efficiency, male: female ratio, normal karyotype percentage across treatments but reduced cellular proliferation. To evaluate their physiological status, we are analyzing cellular apoptosis, mitotic cycle, gene expression profiles and metabolic pathways critical in development of a healthy metabolism. These mESCs act as models that mimic the inaccessible embryo and fetus within the mother, providing a mechanistic approach to understand the phenotypic changes induced by environmental effects thereby reducing the total number of animals used. (Funded by BBSRC, EU-FP7, EpiHealth, EpiHealthNet).
11 Jul 2016 - 11 Jul 2016