320 USE OF SYNTHETIC HYALURONAN OR POLYVINYLPYRROLIDONE FOR INTRACYTOPLASMIC SPERM INJECTION INTO MOUSE OOCYTES
Bibliographic record
Abstract
The use of polyvinylpyrrolidane (PVP) in intracytoplasmic sperm injection (ICSI) seems to be exclusively related to its surfactant and colloidal properties. In contrast to PVP, which can be toxic to mouse embryos, hyaluronan (HA) is a biological compound. In addition to its colloidal property, HA plays an important biochemical role in cell proliferation and migration and can be found intracellularly in the cleaving stage of mouse, sheep and primate embryos (Hunter RHF 1994 Mol. Reprod. Dev. 39, 176–181). We expect that the viscoelastic properties of HA in combination with its physiological functions may benefit the ICSI procedure. Oocytes at MII stage were collected from CD-1 mice 14 h after hCG injection (h-pi) and were kept at 37°C in KSOM medium for 30 min before ICSI. Semen used for injection was frozen by direct plunge into liquid nitrogen in M2 medium without cryoprotectants. Samples were thawed at 25°C in the air and mixed (1:5) with M2 medium containing either 10% PVP; 360000 MW (w/v; Sigma, St. Louis, MO, USA) or 60% (v/v) synthetic HA (s-HA; MAP-5; Bioniche Inc, Belleville, ON Canada) with comparable viscosity. Injections were performed at 25°C using a mercury-containing pipette attached to a piezo impact unit (Prime Tech, Ibaraki, Japan). A total of 239 oocytes (115 PVP and 124 s-HA) were injected in groups of ten in four replicates. Individual sperm heads decapitated by the freeze/thaw procedure were injected into oocytes and kept for 15 min at 25°C. Oocytes that survived ICSI were placed in 35 µL drops of KSOM medium (~15 zygotes per drop) under paraffin oil at 37°C and 5% CO2 in humidified air. Cleavage and developmental rates were recorded at 24, 48, and 96 h after oocyte injection. Embryos which developed to the blastocyst stage were transferred to pseudo-pregnant females mated with vasectomized males. At Day 13, recipient mice were sacrificed and the number of implantations and fetuses were recorded. Data were compared between groups by Chi-square analysis. Significantly (P < 0.05) more embryos survived ICSI in PVP (74%) than in s-HA group (56%), which was primarily related to sperm adhesiveness to the injection pipette. However, there were no differences in developmental rates at any stage of in vitro embryo culture between groups (2 cell, 93 vs. 100%; 4–8 cell, 100 vs. 100%; blastocyst, 44 vs 50%) for PVP and s-HA, respectively. Significant differences (P < 0.05) between groups were observed in embryo implantation rates. When ICSI was performed with s-HA, 29 out of 35 blastocysts (83%) transferred to synchronized recipients were implanted, which was accomplished only by 19 of the 35 from the PVP group (54%). However, there was no difference between groups in the number of fetuses detected (8 (23%) vs. 9 (26%) for PVP and s-HA, respectively). The use of s-HA for mouse ICSI can be a valuable alternative to PVP. Hyaluronan may show further benefit if sperm adhesiveness to the micropipette can be eliminated, and may be superior to PVP if embryo implantation rates in the s-HA group can be sustained. The authors would like to thank Bioniche, Inc., Belleville, ON, Canada for donating MAP-5.
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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 | 0.001 | 0.000 |
| Meta-epidemiology (narrow) | 0.001 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.001 |
| Bibliometrics | 0.001 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.001 | 0.000 |
| Research integrity | 0.001 | 0.001 |
| Insufficient payload (model declined to judge) | 0.002 | 0.002 |
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".