global® total® LP for Fertilization H5TF 受精用含蛋白質培養液 (HSA)
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高濃度葡萄糖glucose支持卵母細胞培養和傳統IVF受精
Bicarbonate buffered
Ready-to-use 含 5 mg/ml HSA 蛋白質
|
Product |
Order Code |
Description |
Volume (mL) |
HSA mg/ml |
|
global total LP for Fertilization |
H5TF-010 |
Higher glucose concentration for oocyte culture and conventional fertilization. Bicarbonate buffered |
10 |
5 mg/mL |
Indications for Use
Human oocyte culture and fertilization.
Storage and Shelf Life
Store at 2-8°C and protected from light. Ten (10) weeks from the date of manufacture.
Disposal Consideration
Treat or dispose of waste material in accordance with all local state/provincial, and national requirements. Dispose with laboratory waste.
Composition
Sodium Chloride, Potassium Chloride, Calcium Chloride, Potassium Phosphate, Magnesium Sulfate, Sodium Bicarbonate, Glucose, Lactate Na Salt, Sodium Pyruvate, Glycine, L-Alanine, L-Arginine HCl, L-Asparagine, L-Aspartic Acid, L-Cystine, L-Glutamic Acid, Glycyl-Glutamine, L-Histidine, L-Isoleucine, L-Leucine, L-Lysine HCl, L-Methionine, L-Phenylalanine, L-Proline, L-Serine, L-Threonine, L-Tryptophan, L-Tyrosine, L-Valine, EDTA, Phenol Red, Human Serum Albumin* (5 mg/ml), Gentamicin Sulfate* (10 µg/ml) *from therapeutic-grade source materia
QUALITY CONTROL SPECIFICATIONS
|
Assay (performed for each batch) |
Specification |
|
Physicochemical Tests |
|
|
pH (with 5% CO2) |
7.2-7.4 |
|
Osmolality |
260-270 mOsM |
|
Biological Tests |
|
|
Endotoxin (LAL) |
≤ 0.5 EU/ml |
|
Sterility Test (bacterial and fungal screen, SAL 10–3) |
PASS |
|
Biological Assays |
|
|
1-cell Mouse Embryo Assay (% expanded blastocysts at 96 h of culture) |
≥ 80% |
Special Note on the CO2 Concentration in the Incubator:
In most cases, a 5-7% concentration of CO2 in the incubator will produce a pH of 7.2 to 7.4 in global® total® LP for Fertilization. However, the exact concentration of CO2 required to produce the optimum pH of approximately 7.30 (7.27- 7.33) depends on several factors, including the physical characteristics of incubator and the altitude. Consequently, we strongly recommend that each laboratory determine and use the concentration of CO2 that is required to produce a pH of 7.30 in global® total® LP for Fertilization.
INSTRUCTIONS FOR USE
The procedures described below have been found to be effective for human oocyte culture and fertilization and are offered only as examples. Every laboratory must define and optimize its own procedures.
1. Prepare dishes for oocyte holding and/or fertilization, containing small droplets (25-100 µl) or larger volume (0.5-1.0 ml) of global® total® LP for Fertilization under oil, according to general laboratory practice.
2. Place the culture dishes in the incubator for sufficient time to ensure CO2 and temperature equilibration. Depending on the exact configuration, this may take from 24-48 hours. Equilibration will require less time if the oil and medium have been pre-equilibrated.
3. At the conclusion of the retrieval, dissect the oocytes to remove any degenerate and/or excess cumulus cells, blood and debris, and wash the oocytes, according to your standard laboratory procedures.
4. Transfer the oocytes into the global® total® LP for Fertilization droplets in the oocyte-holding dishes (1-2 oocytes/droplet).
5. Place the oocyte-holding dishes into a CO2 incubator and culture for 3-6 hours.
6. Add sufficient sperm to each droplet of global® total® LP for Fertilization in the fertilization dishes to produce the required sperm concentration.
7. Let the fertilization dishes sit for several minutes and then examine each droplet to ensure that the sperm concentration is appropriate.
8. Transfer the oocytes from the oocyte-holding dishes to the sperm-containing droplets in the fertilization dishes (1-2 oocytes/microdrop).
9. Place the fertilization dishes into a CO2 incubator and culture. 10. The next day, evaluate the oocytes for evidence of fertilization, and wash and transfer them to embryo culture medium, according to your standard laboratory procedures.