Variation in the osmotic characteristics of aging red blood cells: insights for cryopreservation optimization
Bibliographic record
Abstract
Cryopreservation is a long-term storage strategy for maintaining the quality of red blood cells (RBCs) used for clinical and industrial purposes. However, cryopreservation can induce osmotic stress, leading to a 15 to 20% loss of RBCs during freezing/thawing and cryoprotectant addition/removal. This study investigated how the biological aging of RBCs influences their osmotic characteristics. We hypothesized that biologically older RBCs (O-RBCs) would exhibit diminished osmotic features compared to biologically younger RBCs (Y-RBCs), thereby contributing to their loss during cryopreservation. Seven red cell concentrates (RCCs) were pooled, and their density profile was determined using Percoll separation. Y-RBCs and O-RBCs, representing the extremes of the density spectrum, were isolated. Rheological parameters (O hyper , EI max , EI hyper, K EI ) were measured with laser ektacytometry. Osmotic fragility and hemolysis tests were performed to assess subpopulation responses to osmotic stress. Water ( L p ) and solute permeability ( P s ) of these subpopulations, in addition to unseparated RBCs (U-RBCs), were measured using stopped-flow spectroscopy. Aliquots of Y- and O-RBCs were differentially labeled with biotin and spiked back into a pooled unit. The pooled unit was split into five, and cryopreserved via a high glycerol/slow-cooling method, a standard method adopted by international blood banks. Glycerolization and deglycerolization were conducted using the Haemonetics ACP 215 Automated Cell Processor. Flow cytometry was used to assess the recovery of biotinylated RBCs (BioRBCs) before glycerolization and following post-thaw deglycerolization on days 1 and 14. O-RBCs exhibited the highest rigidity (K EI ) and lowest O hyper , EI max , and EI hyper . Osmotic hemolysis and osmotic fragility indicated that O-RBCs exhibited superior tolerance to hypotonic solutions than Y- and U-RBCs ( P < 0.05). O-RBCs demonstrated significantly higher L p values than Y-RBCs across hypo- and hypertonic conditions ( P < 0.05). O-RBCs had the highest P s during deglycerolization ( P < 0.05). Y- and O-BioRBCs showed a stable trend throughout the 14 days of hypothermic storage (1–4°C) post-deglycerolization, with no significant difference between subpopulations. The study revealed that biological aging is associated with alterations in the osmotic and rheological properties of RBCs. Despite notable differences in osmotic characteristics, under the test conditions, survival rates of Y- and O-BioRBCs remained comparable after 14 days of hypothermic storage. These findings potentially support that tailoring cryopreservation protocols to specific RBC subpopulations can improve recovery rates and make cryopreservation more clinically efficient and broadly applicable.
Fetched live from OpenAlex and de-inverted. Abstracts are not stored in this database: the inverted indexes are 8.6 GB of the frame’s 9.3 GB of text, and the host has 13 GB free.
How this classification was reachedexpand
Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.000 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one teacher head, not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".