Investigating Interactions of Trehalose-Containing Liposomes with Human Red Blood Cells
Bibliographic record
Abstract
A major obstacle in using intracellular sugars for the cryopreservation of mammalian cells is the inability of cells to synthesize or actively accumulate these sugars. We are investigating the use of liposomes, which are synthetic, microscopic vesicles, for the intracellular delivery of stabilizing sugars into mammalian cells. This study examines the interactions of trehalose-containing liposomes with human red blood cells (RBCs). Unilamellar liposomes were synthesized using an extrusion method to contain trehalose in the aqueous core. Liposomal preparations were labeled with a lipophilic fluorophore rhodamine B chloride (R18) at quenching concentrations (4 mol%), or the 5(6)-carboxyfluorescein [5(6)-CF] marker to label the intraliposomal aqueous phase. Flow cytometry and fluorescent microscopy were used to assess the interactions between fluorescently labeled liposomes and RBCs. The delivery of liposomal contents into RBCs was assessed by spectrophotometric measurement of intracellular trehalose. The results of this study show that the transfer of fluorescence from liposome to RBC population is due to both liposomal adsorption and fusion. The merger of membranes and lateral diffusion of liposomal lipids into the RBC bilayer resulted in the spatial separation of membrane R18 fluorophores, and therefore, a dequenching effect, marked by a 344 ± 11% increase of the RBC mean fluorescence intensity. The ability of 5(6)-CF and R18 liposomes to deliver their aqueous contents into RBCs was confirmed by intracellular trehalose measurement. After treatment with 5(6)-CF liposomes, RBC trehalose concentration was 0.18 ± 0.03 mM, whereas R18 liposomes delivered 0.96 ± 0.07 mM trehalose. The results from this study strongly support the hypothesis that liposomes can be used as trehalose delivery vesicles. Delineating the mechanism of liposomal interaction with RBCs is an important step toward establishing the use of liposomes as tools for the intracellular delivery of stabilizing disaccharides for biopreservation purposes.
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| 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) | 0.001 | 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 source (direct Gemma or distilled Codex), 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".