Composition:

• GM501 Gradients consist of silane-coated colloidal silica particles suspended in HEPES-buffered EBSS (Earle’s balanced salt solution).

Produktspezifikationen und Qualitätskontrolle:

• All raw materials are of highest available purity (European Pharmacopoeia and/or USP standard) if applicable.
• A certificate of analysis is available for each batch upon request from our website with respective lot number.
• The MSDS for GM501 Gradient 45% and 90% is available upon request and can also be downloaded from our website.
• GM501 Gradient 45% and 90% is manufactured according to the following specifications:

• The intended users are IVF professionals (lab technicians, embryologists or medical doctors).

Calculation of g-forces:

• The g-force of your centrifuge can be calculated using this formula:

g = 1.118 x r x rpm2 or rpm = Square root {g/ (1.118 x r)}
r = radius of centrifuge in mm
rpm = rotations per minute/1000

Example 1
r = 150 mm
rpm = 1200 rotations per minute
g = 1.118 x 150 x 1.44 = 242 g
Example 2
r = 150 mm
g = 300 g
rpm = SQR {300/ (1.118 x 150)} = 1.33
rpm = 1330 rotations per minute

Instructions for use:

• Importantly: each laboratory should consult its own optimized and validated laboratory procedures.

Instruction for use with fresh semen samples:

Mix the density gradient bottles by 5 bottle inversions before use.

1. Bring all components of the system and samples to room temperature or to 37°C.
2. Transfer 2.5 ml of Gradient 45% into a sterile disposable centrifuge tube.
3. Using a 3cc syringe with a 1 1/2” 21 g needle, place 2.5 ml of Gradient 90% under the Gradient 45%. Take care that the two layers are distinctly separated. This is done by placing the tip of the needle on the bottom of the test tube and slowly dispensing the Gradient 90%. This two-layer gradient is stable for up to two hours.
4. Gently place up to 2.5 ml of liquefied semen onto the Gradient 45% using a transfer pipette or syringe.
5. Centrifuge for 15 to 18 minutes at 350 g to 400 g. When this centrifugation is completed, you may not be able to visibly see a pellet. If so, it is essential to continue the procedure with a second centrifugation of 3 to 5 minutes.
6. Remove supernatant down to the pellet.
7. Using a syringe, add 2-3 ml of sperm washing medium and resuspend the pellet.
8. Centrifuge for 8 to 10 minutes at 300 g.
9. Higher sperm concentration will require the maximum 10 minutes centrifugation to ensure a complete and thorough sperm wash.
10. Remove supernatant down to the pellet and repeat steps 7 and 8.
11. Remove supernatant and replace with a suitable volume of appropriate medium.

Instructions for use with frozen semen samples:
Mix the density gradient bottles by 5 bottle inversions before use.

1. Bring all components of the system and samples to room temperature or to 37°C.
2. Transfer 1 ml of Gradient 45% into a sterile disposable centrifuge tube.
3. Using a 3cc syringe with a 1 1/2” 21 g needle, place 1 ml of Gradient 90% under the Gradient 45%. Take care that the two layers are distinctly separated. This is done by placing the tip of the needle on the bottom of the test tube and slowly dispensing the Gradient 90%. This two-layer gradient is stable for up to two hours.
4. Gently place the thawed semen sample onto the Gradient 45% using a transfer pipette or syringe (0.5 ml maximum).
5. Centrifuge for 15-20 minutes at 350 g.
6. Remove supernatant down to (but not below) the 0.5 ml mark above the pellet.
7. Using a syringe, add 2-3 ml of sperm washing medium and resuspend the pellet.
8. Centrifuge for 8 to 10 minutes at 300 g.
9. Remove supernatant down to the pellet and repeat steps 7 and 8.
10. Remove supernatant and replace with a suitable volume of appropriate medium.

If samples do not liquefy and therefore do not pass through the layers, increasing the centrifugal force up to, but no more than, 500 g will help to separate the sperm.

Precautions and warnings:

• All human, organic material should be considered potentially infectious. Handle all specimens as if capable of transmitting HIV or hepatitis.
• Always wear protective clothing when handling specimens.
• Aseptic technique must be used to avoid possible contamination.
• Any serious incident (as defined in European Medical Device Regulation 2017/745) that has occurred must be reported to Gynemed and, if applicable, the Competent Authority of the EU Member State in which the user and/or patient is established.
• Only for the intended use.

Warning before use:

• Do not use the product if it becomes discolored (medium contains phenol red), cloudy, or shows any evidence of microbial contamination.
• Do not use the product if seal of the container is opened or defective when the product is delivered.
• Do not use the product if the expiry date has been exceeded.
• Do not freeze before use.
• Do not re-sterilize after opening.
• Depending on the number of procedures that will be performed on one day, remove the required volume of medium under aseptic conditions in an appropriate sterile recipient. This is in order to avoid multiple openings/warming cycles of the medium. Discard excess (unused) media.

Storage instructions and stability:

• The shelf life is 18 months from time of manufacture.
• Store products between 2-25°C before first use, once opened at 2-8°C.
• The product can be used safely up to 7 days after opening, if sterile conditions are maintained, and the products are stored at 2-8°C.
• Stable after transport at elevated temperatures (up to 5 days at ≤ 37°C)
• The devices need to be disposed in accordance with local regulations for disposal of medical devices.
• Keep away from (sun)light