Biopreservation and Biobanking最新文献

In this study, the effects of ferulic acid (0.1, 1, ve 10 mM), tryptophan (5, 25, ve 50 mM), and L-glutamine (10, 50, ve 100 mM) at different doses added to 18% raffinose + 3% skimmed milk powder sperm extender on the freezing of mouse spermatozoa in liquid nitrogen were investigated. The combination of 18% raffinose + 3% skimmed milk powder without additives was used as the control group. Frozen spermatozoa were thawed in a 37°C water bath for 30 seconds. After freeze-thawing, motility, dead spermatozoa ratio, plasma membrane integrity, abnormal acrosome ratio, motility endurance (for 4 hours), and cell apoptosis tests were performed in Human Tubal Fluid (HTF). Compared with the control group after freezing and thawing, the highest motility and plasma membrane integrity were obtained in the 10 mM L-glutamine group with 56.6% ± 2.11% and 77.8% ± 0.87%, respectively (p < 0.05). In addition, when compared to the control group, the lowest rate of dead spermatozoa and abnormal acrosome was found in the 10 mM L-glutamine group as 26.0% ± 1.46% and 6.3% ± 1.09%, respectively (p < 0.05). The highest motility values for spermatozoa endurance were determined in the 10 and 50 mM L-glutamine groups up to the 4th hour compared to the control group (p < 0.05). In the evaluation of apoptosis in semen samples, there was no significant difference between the control, 0.1 mM ferulic acid, and 10 mM L-glutamine groups (p > 0.05). As a result, it was determined that the addition of 10 mM L-glutamine to the spermatozoa extender increased the motility, viable spermatozoa, functional membrane integrity, intact acrosome ratios, or motility endurance after freeze-thawing and could be used successfully in the freezing extender of mouse spermatozoa.

Development of novel biomarkers for diagnosis of disease and assessment of treatment efficacy utilizes a wide range of biospecimens for discovery research. The fitness of biospecimens for the purpose of biomarker development depends on the clinical characteristics of the donor and on a number of critical and potentially uncontrolled pre-analytical variables. Pre-analytical factors influence the reliability of the biomarkers to be analyzed and can seriously impact analytic outcomes. Sample quality stratification assays and tools can be utilized by biorepositories to minimize bias resulting from samples' inconsistent quality. In this study, we evaluated the quality of biobanked specimens by comparing analytical outcomes at 1, 5, and 10 years after collection. Our results demonstrate that currently available assays and tools can be used by biobank laboratories to support objective biospecimen qualification. We have established a workflow to monitor the quality of different types of biospecimens and, in this study, present the results of a qualification exercise applied to fluid samples and their derivatives in the context of urological diseases.

This study examined the influence of heat exposure on DNA samples during polymerase chain reaction (PCR) detection. In this study, λDNA samples, as model DNA, were exposed to 105°C for 3-90 minutes or to 105°C-115°C for 15 minutes by autoclaving. The exposed samples were subjected to real-time PCR using nine primer sets with amplicon sizes of 45-504 bp. Regarding DNA samples exposed to 105°C by autoclaving, the data showed negative correlations between the logarithm of λDNA concentration (log λDNA) calculated using real-time PCR and exposure duration and a good relationship between the slope of the regression line and amplicon size. Regarding λDNA samples exposed to heat for 15 minutes, the data showed negative correlations between the log λDNA and exposure temperature and a good relationship between the slope of the regression line and amplicon size. These results showed that the equations used in this study could predict the degree of degradation in λDNA samples by autoclaving, and the PCR detection levels of the DNA at each amplicon size.

Cell culture is a critical platform for numerous research and industrial processes. However, methods for transporting cells are largely limited to cryopreservation, which is logistically challenging, requires the use of potentially cytotoxic cryopreservatives, and can result in poor cell recovery. Development of a transport media that can be used at ambient temperatures would alleviate these issues. In this study, we describe a novel transportation medium for mammalian cells. Five commonly used cell lines, (HEK293, CHO, HepG2, K562, and Jurkat) were successfully shipped and stored for a minimum of 72 hours and up to 96 hours at ambient temperature, after which, cells were recovered into standard culture conditions. Viability (%) and cell numbers, were examined, before, following the transport/storage period and following the recovery period. In all experiments, cell numbers returned to pretransport/storage concentration within 24-48 hours recovery. Imaging data indicated that HepG2 cells were fully adherent and had established typical growth morphology following 48 hours recovery, which was not seen in cells recovered from cryopreservation. Following recovery, Jurkat cells that had been subjected to a 96 hours transport/storage period, demonstrated a 1.93-fold increase compared with the starting cell number with >95% cell viability. We conclude that CellShip^® may represent a viable method for the transportation of mammalian cells for multiple downstream applications in the Life Sciences research sector.

Semen preservation is a significant biotechnology used to safeguard the genetic material of birds, especially those with declining populations, through biobanking. However, there are limited reports on the successful chilling or cryopreservation of wild bird semen. In general, these techniques are not yet well-established for several species of wild birds and pose several challenges such as the need for bird handling and training, contamination of semen samples, low volume of semen collected, and inefficient preservation protocols. To address these challenges and improve post-thawing outcomes, new possibilities are being investigated, including alternative collection methods to traditional digital massage, the use of antioxidants and enzymes in the medium for chilling or freezing, storage methods using different straws from the usual pellet, and slower freezing rates. This review aims to discuss the various aspects of applying semen preservation in wild birds to create germplasm banks, highlighting the primary results obtained and the challenges that need to be addressed.

Research biobanks have become crucial collaborators in a variety of basic and clinical research projects with comprehensive biological sample collection and associated data storage. Medical students, who are the most important stakeholders of biobanks as future physicians, need to be trained in biobanking; however, there is no consensus on how to include it in formal education. This study aimed to determine and increase awareness among medical students regarding biobanks through peer training organized online by medical student research association networks. Volunteer medical or graduate students were trained by biobank professionals at the Izmir Biomedicine and Genome Center (IBG) biobank for 6-9 months. Then, a biobank event was planned by these trainees, the Ege Scientific Research Team (ESRT), and IBG-Biobank with the support of The Biobanking and BioMolecular Resources Research Infrastructure (BBMRI) Turkey. The study reached students of 46 different medical faculties. Before the event, students' level of knowledge about biobanks was identified using a pre-event questionnaire (n = 239). Following 2 days (4 main sessions) of online events, a post-event questionnaire was administered to event participants (n = 110) and 80.9% of them answered (n = 89). The pre-event survey revealed that only 34.3% of the medical students had heard of the term "Biobank" in Turkey. After the event, medical students were significantly more enthusiastic about putting effort into biobanking and using and sharing stored biobank samples of their patients compared with the pre-event (p < 0.0001). Moreover, 92% of the students stated that they would consider attending an advanced course in biobanking. In conclusion, the current study demonstrates that extracurricular courses with peer learning methods coordinated with medical student associations can be valuable in increasing future physicians' awareness and knowledge of biobanking.

The genetic diversity of endangered deer species, such as Mazama jucunda, can be preserved with the help of somatic cell cryopreservation. This procedure allows obtaining several cells from the individual even after its death, which is very important for applications in reproductive biotechnologies. This study's objective was to test cryopreservation protocols of skin fragments of M. jucunda, using different cryoprotectants in slow freezing. We evaluated four treatments, composed of three cryoprotectants, dimethyl sulfoxide (DMSO), polyvinylpyrrolidone (PVP), and ethylene glycol (EG), used alone and in combination. There was also a control group where the tissue did not undergo cryopreservation. Skin fragments were collected from the medial region of the pelvic limbs of three individuals. Each fragment was divided into 10 equal parts, standardized by weight, making two pieces for each treatment and control from each animal. The collected fragments were evaluated in culture, based on the speed of occupation of the free spaces of the cell culture flask. Cell viability was also evaluated using Trypan Blue dye and the mitotic index to understand the effect of toxicity and freezing on cell membrane integrity and cell division capacity, respectively. The treatments that used association with PVP proved to be more damaging to the cells, taking longer to reach confluence. EG alone showed better results than DMSO in the slow-freezing protocol. Clinical Trial Registration Number is 1390/21.