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Anatomy is taught in the early years of an undergraduate medical curriculum. The subject is volatile and of voluminous content, given the complex nature of the human body. Students frequently face learning constraints in these fledgling years of medical education, often resulting in a spiraling dwindling academic performance. Hence, there have been continued efforts directed at developing new curricula and incorporating new methods of teaching, learning and assessment that are aimed at logical learning and long-term retention of anatomical knowledge, which is a mainstay of all medical practice. In recent years, artificial intelligence (AI) has gained in popularity. AI uses machine learning models to store, compute, analyze and even augment huge amounts of data to be retrieved when needed, while simultaneously the machine itself can be programmed for deep learning, improving its own efficiency through complex neural networks. There are numerous specific benefits to incorporating AI in education, which include in-depth learning, storage of large electronic data, teaching from remote locations, engagement of fewer personnel in teaching, quick feedback from responders, innovative assessment methods and user-friendly alternatives. AI has long been a part of medical diagnostics and treatment planning. Extensive literature is available on uses of AI in clinical settings, e.g., in Radiology, but to the best of our knowledge there is a paucity of published data on AI used for teaching, learning and assessment in anatomy. In the present review, we highlight recent novel and advanced AI techniques such as Artificial Neural Networks (ANN), or more complex Convoluted Neural Networks (CNN) and Bayesian U-Net, which are used for teaching anatomy. We also address the main advantages and limitations of the use of AI in medical education and lessons learnt from AI application during the COVID-19 pandemic. In the future, studies with AI in anatomy education could be advantageous for both students to develop professional expertise and for instructors to develop improved teaching methods for this vast and complex subject, especially with the increasing paucity of cadavers in many medical schools. We also suggest some novel examples of how AI could be incorporated to deliver augmented reality experiences, especially with reference to complex regions in the human body, such as neural pathways in the brain, complex developmental processes in the embryo or in complicated miniature regions such as the middle and inner ear. AI can change the face of assessment techniques and broaden their dimensions to suit individual learners.

Introduction: Many countries adopted remote teaching upon the declaration of COVID-19 pandemic. Like many universities around the world, Sultan Qaboos University in the Sultanate of Oman shifted to emergency remote teaching (ERT) to contain the spread of the virus. In addition, anatomy classes were shifted to be taught online completely. This study aimed to explore students' perspective about the impact of COVID-19 pandemic on anatomy learning. Methodology: Students enrolled in anatomy courses during spring 2020 semester were asked to fill a questionnaire within 4 weeks of the end of online classes. The questionnaire included questions about students' perceptions and preferences about learning anatomy during ERT. Results: Findings indicated that coronavirus pandemic was disruptive to students' anatomy learning. Fulfilling courses requirements, home environment, internet connection, the anatomy knowledge without exposure to cadavers, and other resources in the lab were their main concerns. The reliability of internet connection was a major factor affecting their way of studying anatomy and selecting learning resources. Social media platforms were helpful in assisting students in their study and communication between themselves and their instructors. However, assessment and practical part teaching were not satisfactory to the students and they urge improvement. Conclusion: COVID-19 pandemic was disruptive to anatomy education. Internet connection was the major obstacles for the students, while the social media platforms were helpful tools in study and communication. Students urged improvement in the practical part teaching and assessment of the anatomy courses.

In recent times, online teaching and assessment have provided a great opportunity to explore better methods in medical education. There are inconsistent views concerning the effectiveness of online assessment. Hence, the present study aimed to evaluate online teaching and assessment methods in relation to face-to-face methods by comparing students’ performances. The students’ assessment results in two basic anatomy courses, which are part of the Doctor of Medicine and Biomedical Sciences programs at Sultan Qaboos University, were analysed. We compared the students’ mean scores and coefficient of variance in the multiple-choice written exams and the objective structured practical exams during the spring semesters of 2019, 2020, and 2021, containing face-to-face teaching and exams, partial online teaching and online exams, and online teaching and both proctored online and face-to-face exams, respectively. The sudden transition to online teaching and assessment halfway through the semester resulted in higher means and a lower coefficient of variance among students’ scores in both theory and practical exams. However, when the fully adopted online method of teaching and assessment was employed, the mean scores decreased, and the coefficient of variance increased to figures close to those witnessed before the pandemic, when teaching and assessment were face-to-face. This trend applied to both the Doctor of Medicine and Biomedical Sciences programs’ anatomy courses. The results indicate that online assessment of theoretical and practical anatomical knowledge is comparable to that of face-to-face assessment. However, proper planning and preparedness are mandatory to achieve the desired outcomes.

Globally, fertility-related issues affect around 15% of couples. In 20%−30% of cases men are solely responsible, and they contribute in around 50% of all cases. Hence, understanding of in vivo germ-cell specification and exploring different angles of fertility preservation and infertility intervention are considered hot topics nowadays, with special focus on the use of human pluripotent stem cells (hPSCs) as a source of in vitro germ-cell generation. However, the generation of male germ cells from hPSCs can currently be considered challenging, making a judgment on the real perspective of these innovative approaches difficult. Ever since the first spontaneous germ-cell differentiation studies, using human embryonic stem cells, various strategies, including specific co-cultures, gene over-expression, and addition of growth factors, have been applied for human germ-cell derivation. In line with the variety of differentiation methods, the outcomes have ranged from early and migratory primordial germ cells up to post-meiotic spermatids. This variety of culture approaches and cell lines makes comparisons between protocols difficult. Considering the diverse strategies and outcomes, we aim in this mini-review to summarize the literature regarding in vitro derivation of human male germ cells from hPSCs, while keeping a particular focus on the culture methods, growth factors, and cell lines used.

Finding robust culture conditions for in vitro maturation (IVM) of male germ cells is still a challenge. Recently, a testis organ culture method, using Knockout Serum Replacement (KSR), was suggested as a promising approach. However, the efficiency of that model is still not optimal. Hence, we have tried to establish the culture conditions in two laboratories, and to improve the reliability of the culture system to generate mature germ cells. Male mice at three days of age were sacrificed. Testes were cut into small pieces which were cultured atop agarose stands, using Minimum Essential Medium alpha supplemented with different supplements; melatonin, Glutamax, and different concentrations of KSR. The results showed that the duration of culture beyond 18 days had an impact on the number of differentiated germ cells. Supplementation with melatonin and Glutamax revealed a positive influence on the efficiency of male germ cell differentiation in vitro. Furthermore, the results confirmed that KSR had a positive effect on germ cell maturation and testosterone production, with a concentration of at least 10%. In conclusion, this study emphasizes the beneficial role of at least 10% KSR in the IVM of germ cells. Electronic supplementary material The online version of this article (doi:10.1007/s10439-017-1847-z) contains supplementary material, which is available to authorized users.

STUDY QUESTION Does the X chromosome inactivation (XCI) of Klinefelter syndrome (KS)-derived human induced pluripotent stem cells (hiPSCs) correspond to female human pluripotent stem cells (hPSCs) and reflect the KS genotype? SUMMARY ANSWER Our results demonstrate for the first time that KS-derived hiPSCs show similar XCI behavior to female hPSCs in culture and show biological relevance to KS genotype-related clinical features. WHAT IS KNOWN ALREADY So far, assessment of XCI of KS-derived hiPSCs was based on H3K27me3 staining and X-inactive specific transcript gene expression disregarding the at least three XCI states (XaXi with XIST coating, XaXi lacking XIST coating, and XaXe (partially eroded XCI)) that female hPSCs display in culture. STUDY DESIGN, SIZE, DURATION The study used hiPSC lines generated from two azoospermic patients with KS and included two healthy male (HM) and one healthy female donor. PARTICIPANTS/MATERIALS, SETTING, METHODS In this study, we derived hiPSCs by reprograming fibroblasts with episomal plasmids and applying laminin 521 as culture substrate. hiPSCs were characterized by karyotyping, immunocytochemistry, immunohistochemistry, quantitative PCR, teratoma formation, and embryoid body differentiation. XCI and KS hiPSC relevance were assessed by whole genome transcriptomics analysis and immunocytochemistry plus FISH of KS, HM and female fibroblast, and their hiPSC derivatives. MAIN RESULTS AND THE ROLE OF CHANCE Applying whole genome transcriptomics analysis, we could identify differentially expressed genes (DEGs) between KS and HM donors with enrichment in gene ontology terms associated with fertility, cardiovascular development, ossification, and brain development, all associated with KS genotype-related clinical features. Furthermore, XCI analysis based on transcriptomics data, RNA FISH, and H3K27me3 staining revealed variable XCI states of KS hiPSCs similar to female hiPSCs, showing either normal (XaXi) or eroded (XaXe) XCI. KS hiPSCs with normal XCI showed nevertheless upregulated X-linked genes involved in nervous system development as well as synaptic transmission, supporting the potential use of KS-derived hiPSCs as an in vitro model for KS. LIMITATIONS, REASONS FOR CAUTION Detailed clinical information for patients included in this study was not available. Although a correlation between DEGs and the KS genotype could be observed, the biological relevance of these cells has to be confirmed with further experiments. In addition, karyotype analysis for two hiPSC lines was performed at passage 12 but not repeated at a later passage. Nevertheless, since all XCI experiments for those lines were performed between passage 11 and 15 the authors expect no karyotypic changes for those experiments. WIDER IMPLICATIONS OF THE FINDINGS As KS patients have variable clinical phenotypes that are influenced by the grade of aneuploidy, mosaicism, origin of the X chromosome, and XCI ‘escapee’ genes, which vary not only among individuals but also among different tissues within the same individual, differentiated KS hiPSCs could be used for a better understanding of KS pathogenesis. STUDY FUNDING/COMPETING INTEREST(S) This study was supported by grants from the Knut and Alice Wallenberg Foundation (2016.0121 and 2015.0096), Ming Wai Lau Centre for Reparative Medicine (2-343/2016), Ragnar Söderberg Foundation (M67/13), Swedish Research Council (2013-32485-100360-69), the Centre for Innovative Medicine (2–388/2016–40), Kronprinsessan Lovisas Förening För Barnasjukvård/Stiftelsen Axel Tielmans Minnesfond, Samariten Foundation, Jonasson Center at the Royal Institute of Technology, Sweden, and Initial Training Network Marie Curie Program ‘Growsperm’ (EU-FP7-PEOPLE-2013-ITN 603568). The authors declare no conflicts of interest.

In the past, explant tissue-culture methodologies have been used to grow gonads and study their development. Results from in vitro cultures of human gonads showed limited progress toward gonadal cell differentiation and were focused mainly on germ-cell differentiation. Thus, detailed studies focusing on human first-trimester gonadal tissue functionality in vitro are still missing. In this study we investigated the endocrine function of human first-trimester gonads in vitro. We included 27 female and 28 male gonadal samples, derived from a total of 55 cases, at postconceptional ages of 4.5 to 10.5 weeks. Tissues were cultured using an explant tissue-culture system for 14 days. Assays for testosterone (liquid chromatography-tandem mass spectrometry), anti-Mullerian hormone (AMH; ELISA), and inhibin B (ELISA) were performed using media collected after 7 and 14 days of culture. We demonstrated sex- and age-dependent secretion profiles of testosterone, AMH, and inhibin B in the culture media, which resemble the pattern of hormone production in human gonads in vivo, from the few available studies at the same age range. Our study shows that explant tissue-culture conditions are robust for culture of human first-trimester gonadal somatic cells. Thus, it can be used to study human gonadal development and related diseases as well as the effect of potentially hormone-disturbing substances in human gonads during development. However, detailed molecular studies are needed for better understanding of the mechanistic control of the endocrine function of human first-trimester gonads.

Human embryonic stem (hES) cells represent an important tool to study early cell development. The previously described use of human recombinant laminin (LN) 521 represented a step forward in generating clinically safe culture conditions. To test the short-term effect of LN521 on cultured hES cells, five male hES cell lines were cultured on human foreskin fibroblasts (hFFs), Matrigel, LN521, and LN121 and characterized by qPCR, immunofluorescence analysis, as well as their potential for three-germ layer differentiation. Variations in gene expression related to pluripotency, stemness, and testicular cells at different passages and culture conditions were evaluated by qPCR. All cell lines expressed pluripotency markers at protein and RNA level and were able to differentiate into cell types of the three germ layers after being cultured on LN521 for nine passages. Reduction in variation of pluripotency marker expression could be observed after culturing the cells on LN521 for nine passages. hES cells cultured on LN521 exhibited less differentiation, faster cell growth, and attachment when compared to hES cells cultured on LN121 or Matrigel. Our results indicate a positive effect of LN521 in stabilizing pluripotency gene expression and might be the first step towards more controllable and robust culture conditions for hES cells.

The mechanisms underlying human germ cell development are largely unknown, partly due to the scarcity of primordial germ cells and the inaccessibility of the human germline to genetic analysis. Human embryonic stem cells can differentiate to germ cells in vitro and can be genetically modified to study the genetic requirements for germ cell development. Here, we studied NANOS3 and DAZL, which have critical roles in germ cell development in several species, via their over expression in human embryonic stem cells using global transcriptional analysis, in vitro germ cell differentiation, and in vivo germ cell formation assay by xenotransplantation. We found that NANOS3 over expression prolonged pluripotency and delayed differentiation. In addition, we observed a possible connection of NANOS3 with inhibition of apoptosis. For DAZL, our results suggest a post-transcriptional regulation mechanism in hES cells. In addition, we found that DAZL suppressed the translation of OCT4, and affected the transcription of several genes associated with germ cells, cell cycle arrest, and cell migration. Furthermore, DAZL over expressed cells formed spermatogonia-like colonies in a rare instance upon xenotransplantation. These data can be used to further elucidate the role of NANOS3 and DAZL in germ cell development both in vitro and in vivo.