Regulation of self-renewal and detection of karyotypic changes of pluripotent human embryonic stem cells

Human embryonic stem cells are pluripotent cells capable of renewing themselves and differentiating to specialized cell types. Because of their unique regenerative potential, pluripotent cells offer new opportunities for disease modeling, development of regenerative therapies, and treating diseases. Before pluripotent cells can be used in any therapeutic applications, there are numerous challenges to overcome. For instance, the key regulators of pluripotency need to be clarified. In addition, long term culture of pluripotent cells is associated with the accumulation of karyotypic abnormalities, which is a concern regarding the safe use of the cells for therapeutic purposes. The goal of the work presented in this thesis was to identify new factors involved in the maintenance of pluripotency, and to further characterize molecular mechanisms of selected candidate genes. Furthermore, we aimed to set up a new method for analyzing genomic integrity of pluripotent cells. The experimental design applied in this study involved a wide range of molecular biology, genome-wide, and computational techniques to study the pluripotency of stem cells and the functions of the target genes. In collaboration with instrument and reagent company Perkin Elmer, KaryoliteTM BoBsTM was implemented for detecting karyotypic changes of pluripotent cells. Novel genes were identified that are highly and specifically expressed in hES cells. Of these genes, L1TD1 and POLR3G were chosen for further investigation. The results revealed that both of these factors are vital for the maintenance of pluripotency and self-renewal of the hESCs. KaryoliteTM BoBsTM was validated as a novel method to detect karyotypic abnormalities in pluripotent stem cells. The results presented in this thesis offer significant new information on the regulatory networks associated with pluripotency. The results will facilitate in understanding developmental and cancer biology, as well as creating stem cell based applications. KaryoliteTM BoBsTM provides rapid, high-throughput, and cost-efficient tool for screening of human pluripotent cell cultures.

Designing new learning experiences? : exploring corporate E-learners’ self-regulated learning

The context of this study is corporate e-learning, with an explicit focus on how digital learning design can facilitate self-regulated learning (SRL). The field of e-learning is growing rapidly. An increasing number of corporations use digital technology and elearning for training their work force and customers. E-learning may offer economic benefits, as well as opportunities for interaction and communication that traditional teaching cannot provide. However, the evolving variety of digital learning contexts makes new demands on learners, requiring them to develop strategies to adapt and cope with novel learning tools. This study derives from the need to learn more about learning experiences in digital contexts in order to be able to design these properly for learning. The research question targets how the design of an e-learning course influences participants’ self-regulated learning actions and intentions. SRL involves learners’ ability to exercise agency in their learning. Micro-level SRL processes were targeted by exploring behaviour, cognition, and affect/motivation in relation to the design of the digital context. Two iterations of an e-learning course were tested on two groups of participants (N=17). However, the exploration of SRL extends beyond the educational design research perspective of comparing the effects of the changes to the course designs. The study was conducted in a laboratory with each participant individually. Multiple types of data were collected. However, the results presented in this thesis are based on screen observations (including eye tracking) and video-stimulated recall interviews. These data were integrated in order to achieve a broad perspective on SRL. The most essential change evident in the second course iteration was the addition of feedback during practice and the final test. Without feedback on actions there was an observable difference between those who were instruction-directed and those who were self-directed in manipulating the context and, thus, persisted whenever faced with problems. In the second course iteration, including the feedback, this kind of difference was not found. Feedback provided the tipping point for participants to regulate their learning by identifying their knowledge gaps and to explore the learning context in a targeted manner. Furthermore, the course content was consistently seen from a pragmatic perspective, which influenced the participants’ choice of actions, showing that real life relevance is an important need of corporate learners. This also relates to assessment and the consideration of its purpose in relation to participants’ work situation. The rigidity of the multiple choice questions, focusing on the memorisation of details, influenced the participants to adapt to an approach for surface learning. It also caused frustration in cases where the participants’ epistemic beliefs were incompatible with this kind of assessment style. Triggers of positive and negative emotions could be categorized into four levels: personal factors, instructional design of content, interface design of context, and technical solution. In summary, the key design choices for creating a positive learning experience involve feedback, flexibility, functionality, fun, and freedom. The design of the context impacts regulation of behaviour, cognition, as well as affect and motivation. The learners’ awareness of these areas of regulation in relation to learning in a specific context is their ability for design-based epistemic metareflection. I describe this metareflection as knowing how to manipulate the context behaviourally for maximum learning, being metacognitively aware of one’s learning process, and being aware of how emotions can be regulated to maintain volitional control of the learning situation. Attention needs to be paid to how the design of a digital learning context supports learners’ metareflective development as digital learners. Every digital context has its own affordances and constraints, which influence the possibilities for micro-level SRL processes. Empowering learners in developing their ability for design-based epistemic metareflection is, therefore, essential for building their digital literacy in relation to these affordances and constraints. It was evident that the implementation of e-learning in the workplace is not unproblematic and needs new ways of thinking about learning and how we create learning spaces. Digital contexts bring a new culture of learning that demands attitude change in how we value knowledge, measure it, define who owns it, and who creates it. Based on the results, I argue that digital solutions for corporate learning ought to be built as an integrated system that facilitates socio-cultural connectivism within the corporation. The focus needs to shift from designing static e-learning material to managing networks of social meaning negotiation as part of a holistic corporate learning ecology.