Gradient Medium
  • Gradient Medium

Gradient Medium

Gradient Medium

Application: Washing and preparing human sperm for use in ART processes

REF 45%: RS-GM4550 50 ml
REF 90%: RS-GM9050 50 ml
REF 100%: RS-GM10050 50 ml

Gradient environment brochure (download)

Gradient environment license (download)

Gradient environment sterility test report (download)

The report of the in vitro fever generation test of the gradient medium (download)

report of fever generation test by LAL gradient medium method (download)

Gradient environmental stimulation test report (download)

Gradient environment sensitization test report (download)

Genotoxicity test report of gradient medium (download)

The cytotoxicity test report of the gradient medium (download)

Lifetime test report of the gradient environment packaging system (download)

General information

Application: Washing and preparing human sperm for use in ART processes

Storage conditions and duration: Store in a refrigerator at 2-8°C and away from light. This product can be used up to 24 months after production (if the sterile packaging is not opened).

Considerations for waste disposal: It should be done according to the laboratory waste disposal instructions.

Ingredients: This product contains bicarbonate buffer and HEPES, glucose, lactate and pyruvate needed to meet the nutritional needs of sperm in laboratory conditions outside the CO2 incubator.


The efficiency of the following protocol for washing and preparing sperm with gradient medium has been shown, and this protocol is presented only as an example:

1- To create concentrations of 45 and 90% of the gradient medium, the gradient medium 0 should be mixed with the sperm washing medium.
2- The required volume of 45% and 90% concentrations of the gradient medium is removed and placed in a CO2-free incubator for a sufficient period of time (1-2 hours) to ensure the establishment of temperature balance. In case of using a CO2 incubator, in order to prevent pH drop, make sure that the lid of the container containing the media is completely closed.
3- Slowly adding 1.5 ml of 45% gradient medium to the tube containing 1.5 ml of 90% gradient medium (the 45% gradient medium should be placed above the 90% gradient medium).
4- Slowly add 1 ml of semen to the above tube and centrifuge the tube for 20 minutes at 400-150 g.
B Draining and removing the remains of seminal fluid and 45% and 90% gradient environments.
6- Collect the formed sediment with the minimum possible amount from the supernatant and transfer it to a sterile tube containing 5 ml of the sperm washing medium.
7- Washing twice with a centrifuge for 10 minutes at 400-150 g.
8- Washing twice with a centrifuge for 10 minutes at 400-150 g.


1- The user must read this manual completely and receive the necessary training before using the product.
2- This product cannot be used for injection into the body.
3- Do not re-sterilize the product.
4- Avoid using the product if you see the following:
- Presence of damage in sterile packaging
- The expiration date has passed
- Color change, turbidity or the presence of suspended particles or sediment.
5- This medium contains small amounts of sodium bicarbonate buffer and therefore does not need to be exposed to CO2 to adjust the pH before or during use. Because this environment contains HEPES buffer, its exposure to CO2 causes toxicity.
6- When using this product, in order to prevent contamination, aseptic methods should be used.
7- This product can be used safely up to 7 days after opening the sterile packaging.