Biopreservation and Biobanking

Biological material and health information from patients are valuable for medical research. Under a “broad” consent model, hospital patients in Norway can consent to their biological material and health information being stored in research biobanks and used for “specific, broadly defined research purposes” within a specified medical research area but not for medical research in general. Patients are asked to provide new consent each time researchers wish to use their material in a different medical research area. This study investigated patient representatives’ views on having a general consent for medical research without limitation to specific research purposes. We also investigated preferences for the storage of biological samples, the process of consent collection, and factors motivating or hindering consent.

An online, anonymous survey was shared with patient representatives from patient advisory councils at hospitals in Norway, who answered the survey on behalf of patients. A total of 157 representatives completed the survey (response rate of 41%).

A majority (66.2%) supported general consent for medical research and the use of surplus material for medical research in general (63.7%) without limitation to specific research purposes. A minority (35%) supported the use of surplus material without being informed. Sixty-five percent agreed that biological samples could be stored with no time limitation. Over half (56%) preferred to ask patients to consent prior to a hospital visit, and the majority (70.7%) supported the possibility of choosing between digital or paper consent. Factors motivating consent included the desire to contribute to medical research (89.8%) and faith in scientific progress (24.2%). Main hindrances included the fear that health information may be used for other purposes than research (49%), uncertainty regarding research uses (43.9%), and lack of information (31.8%).

A move toward a general consent for medical research may better comply with patients’ wishes and maximize research potential.

Rare disease research in South Africa (SA) faces significant challenges, including limited prioritization and awareness, which hinder advancements in patient care and scientific discovery. This article explores the recruitment strategies employed by the Centre for Human Metabolomics (CHM) Biobank, a nonhospital-based academic rare disease biobank, to address these challenges.

We explain the consent process and documents as well as the three recruitment models employed, namely (1) Recruitment via referring clinician, (2) implementation of monthly diagnostic follow-up sessions, and (3) recruitment of patients for specific projects through clinic-based recruitment drives.

We discuss the benefits as well as the challenges of each model. Challenges included clinician and patient time constraints, distrust from current consent practices, and limited public awareness. We elaborate on future strategies to address these gaps such as simplifying consent, expanding recruitment sites, collaborating with clinical, academic and public institutions, and raising public awareness of the role of the CHM Biobank.

From the models employed over the past 5 years, it is evident that recruitment is most effective when patients perceive a direct benefit, such as involvement in active projects. These strategies outlined in the discussion are crucial for ensuring the CHM Biobank’s sustainability, diversity, and its impact on scientific research and patient outcomes in SA.

Biobanking of prostate cancer tissue is crucial for advancing biomarker-guided precision medicine. However, there is no standardized optimal protocol for biobanking prostatectomy specimens. This study aims to compare the representativeness and sustainability of two biobanking protocols—“Punch” and “Slice”—currently used in Norway.

Fresh frozen tissue from 40 radical prostatectomy specimens was biobanked using both the Punch and Slice protocols. Following macroscopic evaluation, a 2 mm thick transverse slice of the prostate (Slice protocol) was collected and stored in an ultra-freezer for future drill biopsy subsampling, guided by histopathological assessment of adjacent formalin-fixed, paraffin-embedded tissue sections. After the slice was collected, five cylindrical tissue samples were punched from the cut surfaces (Punch protocol). Statistical analyses were conducted to compare the sampling precision and time consumption of both protocols.

Cancerous tissue was successfully sampled in 87.5% of cases using the Punch protocol and 75% of cases using the Slice protocol. Both methods yielded comparable results in terms of the number of cancerous cores and the ability to sample tissue representing the highest Gleason grade. The mean biobanking time of tissue slices was 4.9 minutes compared to 15.1 minutes for the ready-to-use tissue punches. Both methods have previously been shown to provide high-quality RNA extracts.

Both biobanking protocols are effective for sampling prostate cancer tissue, with no significant difference in precision or quality. The choice between protocols should consider factors such as resource availability, tissue quantity, and specific research needs. The Punch protocol is less resource-intensive overall, while the Slice protocol collects vastly more tissue, has a shorter period of ischemia, and provides detailed mapping of biobanked components, allowing for further subsampling at multiple time points.

Human mesenchymal stromal cells (MSCs) are attractive for both medical practice and biomedical research. Nonfreezing short-term storage may provide safe and simple transportation and promote the practical use of MSCs.

We aimed to determine the duration of efficient storage at ambient temperature (22°C) of human dermal MSCs in different three-dimensional organization and to investigate the role of cell metabolic mode in the resistance to the ambient storage damaging factors.

MSCs in monolayer, suspension, and encapsulated in alginate microspheres (AMS) were stored in sealed containers at 22°С in culture medium. Viability (fluorescein diacetate /ethidium bromide) and metabolic activity (Alamar Blue assay) were assessed at 0, 3, 7, 10, and 14 days of the storage. Mitochondrial membrane potential (JC-1 test), cell cycle analysis, reactive oxygen species level, and resistance to hydrogen peroxide were analyzed under culture conditions.

Alginate encapsulation was shown to maintain viability (about 85%), metabolic activity, and adhesion ability during storage for 7 days. The storage of MSCs in both monolayer and suspension was less efficient. Culture of MSCs in AMS decreased basal metabolic activity, mitochondrial activity, and led to reversible cell cycle arrest compared to standard two-dimensional culture. MSCs in AMS have a lower basal level of reactive oxygen species and higher resistance to hydrogen peroxide compared with those in monolayer culture.

Revealed shift into quiescent metabolic mode is essential for alginate-encapsulated MSCs resistance to storage at ambient temperature.

Personalized medicine emphasizes prevention and early diagnosis by developing genetic screening and biomarker assessment tools. Biobanks, including University of Piemonte Orientale (UPO) Biobank, support this effort by providing high-quality biological samples collected, processed, and stored using optimized standardized protocols. To determine the optimal long-term storage conditions for biospecimens used in biomedical research, we evaluated plasma and serum samples cryopreserved using two storage methods, cryovials and straws, across various analytical methodologies with differing sensitivity and robustness.

Plasma and serum samples cryopreserved in liquid nitrogen in vials and straw at the UPO Biobank were subjected to multiple analyses including standard biochemical laboratory analysis, targeted lipidomics, untargeted proteomics, and targeted metabolites quantification through mass spectrometry-based analytical techniques.

Our data demonstrate the robustness and applicability of both storage methods for standard laboratory analyses in evaluating clinically relevant markers in plasma and serum. Lipidomic analysis revealed slight disparities in lipid abundance, though these differences were mostly confined to specific lipid species, particularly fatty acids. Conversely, proteomic and metabolomic analyses uncovered variations in abundance in a significant, albeit limited, fraction of analytes between vials and straw-derived samples.

By highlighting similarities and differences in samples stored in these conditions, this study provides significant insights into optimizing biobanking practices and understanding the factors that influence the integrity of cryopreserved biospecimens and the reliability of the data derived from them. Both straws and vials are convenient and efficient cryopreservation methods, essentially equivalent for samples dedicated to robust and relatively low-sensitive standardized analyses. However, our findings emphasize the need for caution when interpreting omics data from samples subjected to different cryopreservation methods, as subtle variations can arise even with different types of containers.

Ethylene glycol (EG) has been employed as a cryoprotectant for many years in mammalian semen cryopreservation but not assessed for birds except for its recently illustrated beneficial effects on commercial chicken lines. The Indian red jungle fowl is facing trouble in its native range due to human encroachment. Therefore, the present study was designed to elucidate the cryoprotective effect of different EG concentrations (5%, 10%, 15%, and 20%) on frozen Indian red jungle fowl semen.

Semen was collected from 20 cocks, and qualifying ejaculates (>70% motility) were pooled and diluted (15) with red fowl extender. EG was added to the four samples and 20% glycerol in control at 4°C. Samples were equilibrated and cryopreserved in LN₂. Semen quality and biochemical activity were assessed at various stages of cryopreservation.

Sperm motility, viability, plasma membrane and acrosomal integrity, chromatin integrity, and mitochondrial activity were recorded highest (p < 0.05) with 20% EG at the post-equilibration and post-thaw stages. Lipid peroxidation was recorded lowest (p < 0.05) with 20% EG compared with other concentrations and control at the post-equilibration and post-thaw stages.

It is concluded that 20% EG exhibits cryoprotective properties in terms of regulating morphological and biochemical traits of frozen Indian red jungle fowl sperm.