What medium should I use for diluting PureSperm 100?

PureSperm Buffer is the best media to use when diluting PureSperm100. It has been optimised and ionically balanced to best match PureSperm.

Is there any human serum albumin in PureSperm 100?

Our lab research has shown that the yield of motile sperm recovered from the gradient was approximately the same, regardless of whether the gradient layers contained human serum albumin or not. Therefore, we have not deemed it necessary to include any protein in PureSperm 100. The colloid in these products eliminates aggregation of sperm and reduces their sticking to the centrifuge tube. However, human serum albumin must be added to the “wash” solution, as provided in PureSperm Wash.

Are antibiotics included in PureSperm 100?

No, PureSperm 100 does not contain antibiotics. They are not included for the following reasons:

1. The gradient will remove most, if not all, of the bacterial contamination present in the ejaculate, provided that the retrieval of the sperm pellet is carried out according to the instructions given in the package insert.
2. Antibiotics commonly used in cell culture media are toxic to sperm.

What should be used for washing the sperm pellet after the PureSperm 100 gradient centrifugation?

We recommend PureSperm Wash for washing the sperm pellet after gradient centrifugation. The ionic balance of PureSperm Wash lies between that of the gradient and commonly used fertilisation media and will not therefore cause premature hyperactivation of the sperm. PureSperm Wash is also ready-to-use and contains an appropriate level of human serum albumin.

Why do I need to buy a separate product, such as PureSperm Buffer, to dilute PureSperm 100? Could I use PureSperm Wash instead?

You can use PureSperm Wash to dilute PureSperm. However, a comparison in our laboratories has shown that you will obtain a better motile sperm yield by using PureSperm Buffer which has been optimised for this purpose.

Does one need to incubate PS100 and PS40/80 in 5-6% CO², 37°C before use?

The most important thing is to avoid temperature fluctuation in the sperm sample. Ejaculate has a temperature of approximately room temperature or slightly higher. We therefore recommend equilibrating PureSperm to room temperature, no more. The preparation is also processed at room temperature, so this means that an equivalent temperature is maintained.

How do I use PureSperm Buffer?

PureSperm Buffer is used to dilute PureSperm 100 to obtain the layers of different densities required for a discontinuous density gradient. PureSperm Buffer has been carefully formulated to give optimal results when used according to the manufacturer´s instructions. It is ready-to-use, no additives being required.

Why do I need to use PureSperm Buffer to dilute PureSperm 100?

A comparative analysis has shown that you will obtain a better yield of motile sperm by using PureSperm Buffer for diluting PureSperm than by using non-optimised products. Commercially available culture media, washing media, or salt solutions such as Earle’s ´Balanced Salt Solution have not been optimised for this purpose and also require other ingredients need to be added before use, thus increasing the cost.

Why do I need to buy a separate product, such as PureSperm Buffer to dilute PureSperm 100? Could I use PureSperm Wash instead?

Yes, you can use PureSperm Wash to dilute PureSperm 100. However, a comparison in our laboratories has shown that you will obtain a better yield of motile sperm from using PureSperm Buffer which has been optimised for this purpose.

Should I equilibrate PureSperm Buffer before use?

If a bottle of PureSperm Buffer has been previously opened and then stored in the refrigerator, it should be brought up to room temperature before use to avoid cold-shocking the sperm. It is not necessary to equilibrate PureSperm Buffer in the CO² incubator before use.

Are there any antibiotics in PureSperm Buffer ?

No, PureSperm Buffer does not contain antibiotics. Many commonly used antibiotics are toxic to sperm. Furthermore, antibiotics such as penicillin, have a very short half-life in solution. When used according to the manufacturer´s instructions, the PureSperm density gradient will enable an uncontaminated sperm preparation to be obtained from the semen sample.

What tools do I need to perform the sperm preparation using Speedikit?

You only need a centrifuge and some sterile pipettes. PureSperm Speedikit includes the prefilled tubes for both separation and washing of the sample.

Do I need an incubator for performing a sperm preparation with Speedikit?

No, the whole procedure is performed at room temperature.

Can one use the same protocol for SpermCatch as we have for PVP products?

Yes, you can use SpermCatch in the same way as products containing PVP.

What survival rates should we expect after freezing/thawing?

The number of sperm cells surviving a freezing procedure can vary significantly between samples. Analyses on freezing/thawing donor samples however has shown average survival rates of between 65-70%. (“Survival” has been defined as the number of motile sperm cells after thawing compared to the number of motile sperm cells before freezing.)

Can I use Sperm CryoProtec for TESE?

Yes, you can. There is no need to add anything; it’s as simple and easy as using Sperm CryoProtec with ejaculated sperm.

Do I need to add egg-yolk to Sperm CryoProtec before use?

No, you don’t. Sperm CryoProtec is a ready-to-use freezing medium which doesn’t require any additives. Just follow the instructions and your sperm will be able to be safely frozen and thawed.

Does Sperm CryoProtec contain antibiotics?

No, none of our products contains any antibiotics. Adding antibiotics would significantly reduce the shelf life of our products and could “mask” post-sterilisation contamination of the product

Can Blastocysts cultured in any culture media be vitrified with your kits?

VitriBlast /ThermoBlast kit can be used to freeze blastocysts that have been cultured in any culture media.

What device should I use with the VitriBlast kit?

Any device can be used but our general recommendation is to use a closed-system.

Is DMSO more harmful than other cryoprotectants?

No. In general, DMSO is actually one of the least harmful organic solvents. It has a very low toxicity and is not genotoxic, nor mutagenic or carcinogenic.

DMSO is an FDA approved drug used to treat interstitial cystitis, but has been (and is still) widely used to treat various diseases including arthritis, local pains, inflammation, infections (herpes), autoimmune diseases, and gastric ulcer.

In embryology, DMSO was reported to protect membranes better than other cryoprotectants. It is less toxic than propylene glycol and is the standard component of the world’s most successful vitrification mixtures (both human and zoological).