Despite the routine use of cryopreservation for the storage of biological materials, its outcomes are often sub-optimal (including reduced post-thaw viability, recovery, and functionality) due to the damage caused by uncontrolled ice growth. Traditional cryoprotective agents (CPAs), including dimethyl sulfoxide (DMSO), fail to prevent damage caused by ice growth and concerns over CPA cytotoxicity have fostered an increased interest in developing improved CPAs and cryoprotection strategies. The inhibition of ice recrystallization by natural antifreeze (glyco)proteins [AF(G)Ps] to improve cryopreservation outcomes has been examined; however, the ice binding properties of these substances and their challenging large-scale production make them poor CPA candidates. Therefore, the development and deployment of biocompatible, small-molecule ice recrystallization inhibitors (IRIs) for use as CPAs is a worthwhile objective. Extensive structure-activity relationship studies on AF(G)Ps revealed that simple carbohydrate derivatives could inhibit ice recrystallization. It was later discovered that this activity could be fine-tuned by delicately balancing the molecule’s hydrophobicity and hydrophilicity. Current generation small-molecule IRIs have been meticulously designed to avoid binding to the surface of ice and subsequent biological testing (for both cytotoxicity and cryopreservation efficacy) has demonstrated significant improvements to the cryopreservation outcomes of several cell types. However, an individualized cell-specific approach for the simultaneous assessment of multiple cryopreservation outcomes is necessary to realize the full potential of IRIs as CPAs. This article provides a detailed overview of the development of small-molecule carbohydrate-based IRIs and highlights the crucial cell-specific biological considerations that must be taken into account when assessing cryopreservation outcomes.

Uncontrolled ice formation and subsequent growth causes irreversible damage to cells during cryopreservation. Ice recrystallization inhibitors (IRIs) reduce ice crystal growth after nucleation and have proven beneficial for cryopreservation. Controlling ice nucleation can also minimize iceinduced injury and improve the reproducibility of freezing protocols. However, synergy between compounds that influence these processes remains largely unexplored. Herein, we evaluate seven biocompatible carbohydrate-based small-molecules and report that select compounds – 4bromophenyl-β-D-glucose (β-pBrPh-Glc, 1), 4-methoxyphenyl-β-D-glucose (β-PMP-Glc, 2), N-2fluorophenyl-D-gluconamide (2FA, 5), and N-4-chlorophenyl-D-gluconamide (4ClA, 6) – exhibit dual ice-modulating activity, defined as the ability to inhibit ice recrystallization and induce synchronous nucleation, representing a novel mode of ice nucleation control not previously reported. Our results confirm that nucleation control is dependent on carbohydrate structure, where glucose-based molecules are sensitive to AgI-mediated nucleation, producing small ice crystals, whereas galactose-based analogs are insensitive to AgI-mediated nucleation. Overall, our findings reveal that the synergy of ice nucleation and ice recrystallization inhibition is structure- and solvent-dependent.

Objective: Most students spend years in formal education settings without being explicitly taught how to learn effectively. Our objective was to evaluate an innovative intervention designed to effectively equipping all students with learning skills, called the Growth & Goals Module, which is an adaptable open education resource available in English and French.Methods: We evaluated the module using a Practical Participatory Evaluation approach and the 4-level Kirkpatrick Evaluation model. To investigate ten research questions aligned with the model, we collected data from 1845 students and five educators from nine undergraduate courses in science, engineering, and mathematics through questionnaires, focus groups, course assessments, and institutional data.Results: Students and educators reported high satisfaction (Level 1, Learning). The training was new to most students and most completion rates were over 75% when educators provided an incentive. Students in some demographics used the module less than others. In Level 2 (Learning), students’ metacognitive skills increased. They could identify SMART goals and differentiate growth/fixed mindset statements. At Level 3 (Behaviour), students reported intending to use the module in the future. Most educators created learning outcomes for the first time. The module required little time of students and educators; however, greater support, incentives, and rewards are needed for project sustainability. Educators have used the module in courses in many disciplines and levels (Level 4, Results).Conclusions: The Growth & Goals module explicitly teaches core learning skills for students in science, engineering, and mathematics courses and has the potential to scale to other disciplines and levels.

In this dynamic and rapidly changing world, students need to be able to continually learn and adapt throughout their lives. However, most students spend years in formal education settings without being explicitly taught how to learn effectively. To reach our goal of explicitly and effectively equipping all students with learning skills, we developed and evaluated a Growth & Goals module. The module is an Open Education Resource for postsecondary students that educators integrate in their courses to teach core learning skills of metacognition, goal-setting, growth mindset, and mindfulness. Over 5000 students at ten institutions have now used the module. In the present study, we evaluated the module using a Practical Participatory Evaluation approach and the 4-level Kirkpatrick Evaluation model. To answer ten questions aligned with the Kirkpatrick model, we collected data from 1845 students and 5 educators from nine undergraduate courses in science, engineering, and mathematics, which were used to investigate ten research questions aligned with Kirkpatrick’s four evaluation levels. For Level 1 (Reaction), students and educators reported high satisfaction and gave constructive suggestions that centred on expanding the module. The training was new to 88% of students. Most completion rates were over 75% when professors provided an incentive (³ 1%). Students in some demographics used the module less than others: lower-achieving, first-generation university students, from outside the Ottawa-Gatineau area, male, and in certain programs. In Level 2 (Learning), students’ metacognitive skills increased throughout the semester. They could identify SMART goals (Specific, Measurable, Accountable, Reachable, and Time-specific) and differentiate growth/fixed mindset statements. At Level 3 (Behaviour), students applied the module within the originating course, indicated their intent to use the module in the future, and a survey of a subsample indicated that most students used or intended to use the module in a new course. Most educators created course-level learning outcomes for the first time to integrate with the module. As an Open Education Resource with a nearly complete “plug-and-play” format, using the module required little time and low technological skills of students and educators; however, greater support, incentives, and rewards could be provided. Research and development require sustained resources. Finally, in Level 4 (Results), educators have used the module in courses in a number of disciplines, including sciences, engineering, mathematics, education, and psychology. The module addresses institutional goals of transformational learning and agility, as well as two provincial degree level expectations that are rarely explicitly taught in courses. In summary, the Growth & Goals module explicitly teaches core learning skills in a way that is systematic, scalable, and explicit for science, engineering, and mathematics courses, with a potential to expand to any discipline.

Human midbrain organoids (hMOs) are a powerful stem cell-derived model for studying human neurodevelopment and neurological disease in vitro; however, their broad adoption is limited by long culture times, high cost, and inter-batch variability. Cryopreservation offers a strategy to address these limitations by enabling long-term storage and reducing experimental heterogeneity, but its effectiveness depends on robust cryoprotective strategies. Here, we evaluated hMO cryopreservation using a standard 10% dimethyl sulfoxide (DMSO) protocol, alone and in combination with a panel of five ice recrystallization inhibitors (IRIs), including betaine-derived ammonium salts and N-aryl-d-gluconamides. Across all conditions, hMOs were successfully recovered following freeze–thawing, with preservation of overall structural integrity, including maintained morphology and reduced cellular debris. Importantly, post-thaw viability and cell recovery in the 10% DMSO condition were comparable to those of fresh controls, demonstrating that conventional DMSO-based cryopreservation is sufficient to maintain hMO survival and gross structural preservation. While supplementation with select IRIs, particularly N-ethyl betaine ethyl ester iodide (NEBEE-I), yielded modest improvements in viability relative to DMSO alone and a non-significant trend toward enhanced electrophysiological activity in multielectrode array recordings, these effects were secondary to the robust performance of the baseline DMSO protocol. Collectively, these findings establish standard 10% DMSO cryopreservation as an effective and broadly applicable method for hMO preservation, while suggesting that small-molecule IRIs may provide incremental benefits in selected outcome measures.