Mouse embryonic stem cells (mESCs) exist in a naive, primed and ground state of pluripotency. While comparative analyses of these pluripotency states have been reported, the mESCs utilized originated from various genetic backgrounds and were derived in different laboratories. mESC derivation in conventional LIF + serum culture conditions is strain dependent, with different genetic backgrounds potentially affecting subsequent stem cell characteristics. In the present study, we performed a comprehensive characterization of naive, primed and ground state mESCs originating from the same genetic background within our laboratory, by comparing their transcriptional profiles. We showed unique transcriptional profiles for naive, primed and ground state mESCs. While naive and ground state mESCs have more similar but not identical profiles, primed state mESCs show a very distinct profile. We further demonstrate that the differentiation propensity of mESCs to specific germ layers is highly dependent on their respective state of pluripotency.

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STUDY QUESTION Are the transient receptor potential cation channels vanilloid 3 (TRPV3) present and able to mediate strontium (Sr 2+ ) induced artificial activation in human oocytes? SUMMARY ANSWER Sr 2+ did not induce Ca 2+ rises or provoke activation in human oocytes, however, mRNA for the TRPV3 channel was present in metaphase II (MII) human oocytes after IVM and TRPV3 agonists induced Ca 2+ rises and oocyte activation, demonstrating the channels were functional. WHAT IS KNOWN ALREADY Selective activation of TRPV3 by agonists induces Ca 2+ entry and promotes mouse oocyte activation, and the absence of TRPV3 channels in mouse oocytes prevents Sr 2+ mediated artificial activation. Sr 2+ is sometimes used to overcome fertilization failure after ICSI in the clinic, but its efficiency is still controversial and the mechanism(s) of how it mediates the Ca 2+ flux has not been studied yet in human. STUDY DESIGN, SIZE, DURATION The protein distribution ( n = 10) and mRNA expression level ( n = 19) of the TRPV3 channels was investigated in human MII oocytes after IVM. The Sr 2+ (10 mM) and TRPV3 agonists (200 μM 2-aminoethoxydiphenyl borate [2-APB] and 200 μM carvacrol)-induced Ca 2+ response was analyzed in human ( n = 15, n = 16 and n = 16, respectively) and mouse oocytes ( n = 15, n = 19 and n = 26, respectively). The subsequent embryonic developmental potential following the parthenogenetic activation using these three agents was recorded in human ( n = 10, n = 9 and n = 9, respectively) and mouse ( n = 20 per agent) oocytes, by determining pronucleus, or 2-cell and blastocyst formation rates. PARTICIPANTS/MATERIALS, SETTING, METHODS MII oocytes from B6D2F1 mice (6–10 weeks old) as well as human IVM oocytes and IVO oocytes (from patients aged 25–38 years old) with aggregates of smooth endoplasmic reticulum clusters were used. The expression of TRPV3 channels was determined by immunofluorescence staining with confocal microscopy and RT-PCR, and the temporal evolution of intracellular Ca 2+ concentration was measured by time-lapse imaging after exposure to Sr 2+ and TRPV3 agonists (2-APB and carvacrol). Artificial activation efficiency was assessed using these agents. MAIN RESULTS AND THE ROLE OF CHANCE S...

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