Abstract
The use of assisted reproductive technologies (ART) such as in vitro fertilization (IVF) has resulted in the birth of more than 5 million children. While children conceived by these technologies are generally healthy, there is conflicting evidence suggesting an increase in adult-onset complications like glucose intolerance and high blood pressure in IVF children. Animal models indicate similar potential risks. It remains unclear what molecular mechanisms may be operating during in vitro culture to predispose the embryo to these diseases. One of the limitations faced by investigators is the paucity of the material in the preimplantation embryo to test for molecular analysis. To address this problem, we generated mouse embryonic stem cells (mESC) from blastocysts conceived after natural mating (mESCFB) or after IVF, using optimal (KSOM + 5% O2; mESCKAA) and suboptimal (Whitten’s Medium, + 20% O2, mESCWM) conditions. All three groups of embryos showed similar behavior during both derivation and differentiation into their respective mESC lines. Unsupervised hierarchical clustering of microarray data showed that blastocyst culture does not affect the transcriptome of derived mESCs. Transcriptomic changes previously observed in the inner cell mass (ICM) of embryos derived in the same conditions were not present in mESCs, regardless of method of conception or culture medium, suggesting that mESC do not fully maintain a memory of the events occurring prior to their derivation. We conclude that the fertilization method or culture media used to generate blastocysts does not affect differentiation potential, morphology and transcriptome of mESCs.
Highlights
Thirty-six years after the birth of Louis Brown more than 5 million children have been conceived with the use of assisted reproductive technologies (ART) [1]
Mouse embryonic stem cell derivation is not different for in vitro fertilization (IVF) or in vivo blastocysts Blastocysts obtained by flushing the uteri after spontaneous conception (FB) or after in vitro culture (IVFKAA, IVFWM) formed colonies similar in morphology with the same efficiency (Table 1)
To assess the impact of embryo culture on the derivation of embryonic stem cells, we compared the growth of mouse embryonic stem cells (mESC) after derivation over a period of 8 passages (Fig. 1)
Summary
Thirty-six years after the birth of Louis Brown more than 5 million children have been conceived with the use of ARTs [1]. One explanation of how stress during early development may affect long-term health is provided by the developmental origin of health and disease hypothesis (DOHaD) [13]. This theory holds that the embryo or fetus, when exposed to environmental stress, alters its developmental strategy, e.g. gene expression pattern or epigenetic marks, to adapt to the stressful stimulus. The derivation of mouse embryonic stem cells (mESC) from blastocysts would represent a potential solution to this problem. In order to be useful for studies on the mechanism of ART-related changes, embryonic stem cells would need to retain a molecular memory of their original embryo culture environment
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
