global® total® for Fertilization LGTF 受精用培養基 (含蛋白質)

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

A bicarbonate-buffered protein-supplemented medium replete with glucose, lactate, pyruvate and all 20 amino acids is optimal to support the oocyte, attached cumulus cells and sperm.

Sodium Chloride, Sodium Pyruvate, L-Arginine, L-Threonine, Potassium Chloride, L-Alanine, L-Cystine, L-Tryptophan, Calcium Chloride, L-Asparagine, L-Histidine, L-Tyrosine, Potassium Phosphate, L-Aspartic Acid, L-Isoleucine, L-Valine, Magnesium Sulfate, L-Glutamic Acid, L-Leucine, Glycyl-L-Glutamine, Sodium Bicarbonate, Glycine, L-Lysine, EDTA, Glucose, L-Proline, L-Methionine, Phenol Red, Sodium Lactate, L-Serine, L-Phenylalanine, Human Serum Albumin* (4.4 mg/ml), Human α- and β-globulins* (0.6 mg/ml), Gentamicin Sulfate* (10 µg/ml)

*from therapeutic-grade source material

QUALITY CONTROL SPECIFICATIONS

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® 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® 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 appropriate-sized droplets or larger volumes of global® total® 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® for Fertilization droplets in the oocyte-holding dish(es) (1-2 oocytes/ droplet).
5. Evaluate each oocyte according to your standard laboratory protocol.
6. Place the oocyte-holding dish(es) into a CO2 incubator and culture for 3-6 hours; 3-4 hours if the majority of oocytes appear mature, up to 6 hours if the majority appear intermediate or immature.
7. Add sufficient sperm to each droplet of global® total® for Fertilization in the fertilization dishes to produce the required sperm concentration.
8. Let the fertilization dishes sit for several minutes and then examine each droplet to ensure that the sperm concentration is appropriate.
9. Transfer the oocytes from the oocyte-holding dish(es) to the sperm-containing droplets in the fertilization dishes (1-2 oocytes/microdrop).
10. Place the fertilization dishes into a CO2 incubator and culture, according to standard laboratory practice.
11. Evaluate the oocytes for evidence of fertilization, and wash and transfer them to embryo culture medium, according to your standard laboratory procedures.