Biopreservation and Biobanking最新文献

Southeast Asian countries are at the forefront of public health pressures due to a confluence of factors such as population growth, urbanization, environmental pollution, and infectious diseases (re)emergence. Therefore, the ability to be able to conduct research addressing local and regional needs is of paramount importance. As such, biobanking activities, the standardized collection of biological samples, and associated data, developed over the past few decades supporting ongoing biomedical and clinical research, as well as surveillance are of critical importance. However, the regulatory landscape of biobanking is not widely understood and reported, which this narrative review aims to address for the ASEAN member states. It is evident that there are specific regulatory arrangements within each ASEAN member state, which though may be sufficient for the current level of operations, are unlikely to support a regional sharing of biological samples, data, and eventually benefits from the conducted research. Additionally, legacy and often-overlapping regulatory frameworks exist, which raise the need of an eventual consolidation under a single framework. Thus, this field requires further study as well as the creation of viable, practical proposals that would allow for biobanking harmonization and thus the exchange of biological samples and data to be achieved regionally, if not further afield.

Background: Informed consent (IC) for biobank practice is vital to ensure that sample collection, storage, and utilization are ethical. However, the standard practices in biobanking in upper-middle-income countries such as Indonesia often rely on specific consent, leading to restricted sample use and ethical concerns. This article describes the development of an IC model that meets ethical standards and yet is acceptable for biobanking practice in an Indonesian academic hospital. Method: We conducted a study involving Universitas Gadjah Mada (UGM) Biobank Unit and the UGM Academic Hospital, Yogyakarta, Indonesia, between 2019 and 2021. The IC development process consisted of four stages: (1) conceptualization, (2) preparation, (3) pilot, and (4) evaluation. These activities were part of a more extensive pilot study for an academic hospital-based biobank (Medical Biobank for Research in Indonesia (MBRIO) study). Result: We conceptualized a broad consent model, consisting of an information sheet, comprehension test, agreement sheet, and exit survey. We tested and revised the broad consent document to ensure readability, trained 10 consenting staff (1 surgeon and 9 nurses), and then piloted the IC procedure on 24 patients with elective surgery. The evaluation showed that patients understood the information objectively and subjectively. Consenting staff considered the broad consent model acceptable for the academic hospital setting and suggested improvements to increase the readability of information sheets and have more trained staff for better coordination. Conclusion: The IC development process and model consent are ethically sufficient, acceptable and feasible to be implemented in academic hospital-based biobanks in Indonesia adjusted to the business processes.

The purpose of this study, carried out in two experiments, was to investigate the antioxidant effect of Prosopis farcta in Experiment 1, and trehalose in Experiment 2 in the freezing extender, on the quality of frozen-thawed goat epididymal spermatozoa. Sperm samples were added to based egg-yolk Tris-extender containing experimental treatments. The first experimental treatments included the following: an extender of the control group without additive and extender containing 50, 100, or 150 µg/mL of Prosopis farcta ethanolic extract (PEE1, PEE2, and PEE3, respectively). Treatments of the second experiment include an extender of the control group without additive, an extender containing 100 mM of trehalose (Tr), an extender containing 100 µg/mL PEE2, and 100 µg/mL PEE2 + 100 mM Tr. The results of the first experiment showed that PEE2 compared with the control group led to a significant decrease (p < 0.05) in malondialdehyde (MDA) concentration. Also, PEE1 and PEE2 treatments resulted in a significant increase (p < 0.05) in motility parameters by computer-assisted sperm analysis, and MDA concentrations decreased significantly (p < 0.05) in all treatments compared with the control group. In general, the results of the present experiment showed that Prosopis farcta ethanolic extract at the level of 100 µg/mL was effective in improving the quality of frozen-thawed goat epididymal spermatozoa. Also, a combination of Prosopis farcta ethanolic extract and trehalose can be successful in freezing goat epididymal spermatozoa.

We researched the ability of tanacetan pectin from inflorescences of common tansy Tanacetum vulgare L. to change the osmolarity and freezing point of water in solutions of cryoprotectants: glycerol-3.5%, dimethyl sulfoxide (DMSO)-10%, dimethylacetamide-10% (DMAC), and 1.2-propanediol (1.2-PD)-10%, as well as the effect of solutions of tanacetan (0.2%, 0.4%) on the kinetics of crystallization processes and the nature of crystal formation. We used a combination of protector and pectin that we tested earlier, which provided effective protection for human leukocytes and platelets, as well as bovine spermatozoa, at temperatures below freezing (-20°C and -80°C). It has been established that tanacetan slows down the process of water freezing in glycerol, but not in DMSO, DMAC, and 1.2-PD, promotes deeper supercooling of the medium, and affects the morphological structure of ice. The addition of pectin to the cryosolution increases the activity of the main cryoprotectant glycerol even at its low concentrations. The combination of glycerol and tanacetan can be effective in freezing biological materials, which is confirmed by the preservation of leukocytes at -20°C and -80°C for 7 days, platelets at -80°C for 30 days, and spermatozoa at -80°C within 1 day. A comprehensive analysis of the chemical, physicochemical, and cryoprotective properties of tanacetan indicates the prospect of using pectin in the cryopreservation of biological objects at temperatures of electric freezers.

Oocyte vitrification has become a widely adopted method in clinical practice. However, the solidification behavior and its impact on oocytes during the ultrarapid cooling process remain poorly understood. In this study, we established a system and methodology to observe crystallization behavior in oocytes during quench cooling and warming. Subsequently, the threshold concentration of cryoprotective agents (CPAs) required for oocyte vitrification was determined through a visualization method. The results demonstrated that the ice front could not be observed in the image sequence when using 16.5% DMSO +16.5% EG during high-speed quench cooling (2821.58°C/min). Finally, oocytes were encapsulated with an antifreezing hydrogel (7.5% EG +7.5% DMSO +0.5% alginate) and subjected to high-speed quench cooling. No ice crystals appeared in the antifreezing hydrogel-encapsulated oocytes at a low concentration of osmotic CPA (2.4 M). This research opens up new possibilities for oocyte vitrification with a reduced concentration of CPA.

Ozone has been used as a therapy tool in medical science for conditions such as ulcers, peritonitis, wounds, and mostly joint problems. Ozone therapy strengthens the resistance to infections by kick-starting antioxidant, anti-inflammatory, and immune modulation systems. Ozone creates a defensive response against oxidative stress in membranes and protects metabolism against reactive oxygen species (ROS). Sperm membranes are one of ROS's main targets; therefore, the cells' cryopreservation process requires more defensive elements for better results. This study aimed to investigate the protective effect of nano-ozone solution (NOS) on ram sperm cryopreservation and the influence of the process on various sperm parameters for post-thaw (0 hour) and postincubation (6 hours) time points. Samples were collected from six Merino rams in the breeding season by electroejaculation five times at 3-day intervals. The study was conducted by cryopreservation of the samples using a tris citric acid-egg yolk-based extender. The samples were subjected to freezing in control and NOS (0.5, 1, and 2 μg/mL nano-ozone supplemented). Post-thaw motility, hypo-osmotic swelling test, acrosome (fluorescein isothiocyanate-conjugated Pisum sativum agglutinin [PSA-FITC]), and DNA integrities (terminal deoxynucleotidyl transferase dUTP nick end labeling [TUNEL]) were evaluated with a phase-contrast microscope. Mitochondrial membrane potential (MMP) assessments were conducted by JC1-PI dual staining with a flow cytometer. Malondialdehyde and glutathione (GSH) levels were measured by a spectrophotometer. Sperm kinematics were investigated by a computer-assisted sperm analyzer (CASA) at the post-thaw time point. Compared with the control, relatively low doses of NOS (0.5 and 1 μg/mL) yielded better results in many parameters (motility, membrane and acrosomal integrities, MMP, various sperm kinematics, and GSH levels) (p < 0.05). The addition of low ozone doses to cryopreservation extenders improved the results compared with the control group at post-thaw and postincubation time points. Despite the valuable potential of nano-ozone supplementation in ram sperm cryopreservation, this subject requires further investigations with fertility trials soon.

The present study was conducted to evaluate the effects of trehalose supplementation in egg-yolk (EY)-free tris extender on dog spermatozoa. Pooled spermatozoa were diluted with extender 1 (EY-free tris extender supplemented with 0, 10, 15, 20, or 30 mM trehalose) and cooled (2 × 10⁸ sperm/mL) for 1 hour at 4°C. After that, extender 2 (extender 1 containing 1 M glycerol) was added (v:v) to the diluted sperm, loaded in 0.5-mL straws (1 × 10⁸ sperm/mL), and incubated at 4°C for 30 minutes. The sperm straws were frozen over liquid nitrogen (LN₂) vapor for 20 minutes and then plunged directly into LN₂. After thawing at 37°C for 25 seconds, sperm progressive motility (CASA), viability (SYBR-14/PI), apoptosis (Annexin V/PI), and reactive oxygen species (ROS; H₂DCFDA/PI) were evaluated. Thereafter, the optimal concentrations of trehalose were selected, and the gene expression of BAX, BCL2, NOX5, SMOX, OGG1, and ROMO1 was evaluated after freeze-thawing. Supplementation with 20 and 30 mM trehalose significantly increased sperm progressive motility and viability compared to the control. However, trehalose had no significant effect on sperm ROS or phosphatidylserine translocation index. There were minor numerical increases and decreases in gene expression when the selected optimal concentrations of trehalose (20 and 30 mM) were compared to the control. However, there were no significant differences. We conclude that the addition of trehalose (20 and 30 mM) in EY-free extender could improve sperm motility and viability without significant effects on ROS, apoptosis, or gene expression.

There is little guidance concerning biomedical research using tissues from deceased individuals. Unique ethical and legal challenges gained visibility during the coronavirus disease 2019 (COVID-19) pandemic, when important studies using genome sequencing required access to biological materials from deceased individuals. These studies proposed to determine whether specific genomic profiles were associated with important disease outcomes. Such research has previously required consent from next-of-kin or other surrogate decision makers. Ethics waivers for such consent vary within Canada. In Ontario, research ethics boards can grant waivers of consent if the Tri-Council Policy Statement-2 conditions are met. These include that the individual is not harmed, that the materials are essential to the research, and that privacy will be protected. Conversely, in Quebec, Civil Code article 22 imposes an obligation on researchers to seek consent from next-of-kin or another surrogate decision maker with no option for waivers. It became evident to researchers that these standards can sometimes impose an impracticable balance of risks and benefits, especially in public health emergencies. We seek to establish why and when consent requirements should be waived for public health and research involving the tissues of deceased individuals.

Cryopreservation is the most effective technology for the long-term preservation of biological materials, including cells, tissues, and even organs in the future. The process of cooling and rewarming is essential to the successful preservation of biological materials. One of the critical problems in the development of cryopreservation is the optimization of effective rewarming technologies. This article reviewed rewarming methods, including traditional boundary rewarming commonly used for small-volume biological materials and other advanced techniques that could be potentially feasible for organ preservation in the future. The review focused on various rewarming technique principles, typical applications, and their possible limitations for cryopreservation of biological materials. This article introduced nanowarming methods in the progressing optimization and the possible difficulties. The trends of novel rewarming methods were discussed, and suggestions were given for future development.