global® Fertilization LGGF 受精用培養基

Indications for Use

For holding human cumulus-oocyte complexes prior to IVF or ICSI and for conventional IVF.

Storage and Shelf Life

Store at 2-8°C and protected from light. Ten (10) weeks from the date of manufacture.

Composition

For fertilization to occur, a single sperm must pass through the cumulus cells, interact with the zona pellucida, fuse with the oolemma and decondense inside the ooplasm. The fertilization medium must meet the metabolic requirements of the sperm, cumulus cells, and oocyte

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, 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® 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® for Fertilization.

INSTRUCTIONS FOR USE

The procedures described below have been found to be effective for the preparation of global® for Fertilization for holding human cumulus-oocyte complexes prior to conventional fertilization or ICSI, and for conventional in-vitro fertilization. Every laboratory must define and optimize its own procedures.

After each time the original bottle is opened recap the bottle tightly and store at 2-8ºC, protected from light.

Twenty-four (24) hours prior to the use of global® for Fertilization, supplement the medium with either Human Serum Albumin (HSA) or LifeGlobal® Protein Supplement to achieve desired % (v/v) of protein supplementation.

1. Using a sterile pipette or tip, dispense 25-100 µl droplets or in larger volumes (0.5-1.0 ml) of global® for Fertilization supplemented with protein. Cover the oocyte-holding and fertilization microdrops dishes with appropriate oil and place the dishes in a CO2 incubator overnight for gas and temperature equilibration.
2. Before introducing the oocytes, place the dishes in the incubator for a minimum of 8 hours to ensure CO2 and temperature equilibration. Label each dish with patient information.
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. Place the oocyte-holding dishes into a CO2 incubator and culture for 4-6 hours.
4. Transfer oocytes into the supplemented global® for Fertilization microdrops in the oocyte-holding dishes.
5. At time of conventional insemination, add sufficient sperm to each microdrop of supplemented global® for Fertilization in the fertilization dishes to produce the required sperm concentration.
6. Let the fertilization dishes sit for several minutes and then examine each microdrop to ensure that the sperm concentration is appropriate.
7. Transfer the oocytes from the oocyte-holding dishes to the sperm-containing microdrops in the fertilization dishes. Place the fertilization dishes into a CO2 incubator and culture for 15-17 hours (overnight).