The second trimester of human development is marked by asynchronous gonadal development hampering the isolation of homogenous populations of early and late fetal germ cells (FGCs). We evaluated the... Show moreThe second trimester of human development is marked by asynchronous gonadal development hampering the isolation of homogenous populations of early and late fetal germ cells (FGCs). We evaluated the feasibility of using surface markers TNAP, PDPN, EPCAM and ITGA6 to isolate FGCs as well as human primordial germ cell-like cells (hPGCLCs) derived from embryonic stem cells (hESCs) from both sexes by fluorescence-activated cell sorting (FACS). Our results suggest that a combination of TNAP and PDPN was sufficient to separate populations of premeiotic FGCs and hPGCLCs in both sexes. This combination of antibodies also proved efficient in separating 'mitotic' from 'retinoic-acid responsive' female FGCs. Furthermore, we report that the differentiation efficiency of TNAP+PDPN+ hPGCLCs from hESCs was sex-independent, but the ability to propagate differed considerably between the sexes. In contrast to male, female hPGCLCs retained their characteristics and exhibited robust colony-forming ability when cultured for five days in medium containing LIF, forskolin and FGF2. We conclude that marked sex differences exist in the isolation and propagation of human FGCs and hPGCLCs. Our study provides novel insights relevant for the optimization of in vitro gametogenesis in humans. Show less
Responses to surprising events are dynamic. We argue that initial responses are primarily driven by the unexpectedness of the surprising event and reflect an interrupted and surprised state in... Show moreResponses to surprising events are dynamic. We argue that initial responses are primarily driven by the unexpectedness of the surprising event and reflect an interrupted and surprised state in which the outcome does not make sense yet. Later responses, after sense-making, are more likely to incorporate the valence of the outcome itself. To identify initial and later responses to surprising stimuli, we conducted two repetition-change studies and coded the general valence of facial expressions using computerised facial coding and specific facial action using the Facial Action Coding System (FACS). Results partly supported our unfolding logic. The computerised coding showed that initial expressions to positive surprises were less positive than later expressions. Moreover, expressions to positive and negative surprises were initially similar, but after some time differentiated depending on the valence of the event. Importantly, these patterns were particularly pronounced in a subset of facially expressive participants, who also showed facial action in the FACS coding. The FACS data showed that the initial phase was characterised by limited facial action, whereas the later increase in positivity seems to be explained by smiling. Conceptual as well as methodological implications are discussed. Show less