Questions and Answers

• What is the shelf life when the seal has been broken, and SpermFilter® has been aspirated?
  
   

  Ensuring absolute sterile work, SpermFilter^® is stable until the expire date shown on the label. For practical reasons we recommend 1 month shelf life after the seal is broken.
• Is there any clinical documentation available on SpermFilter®
  
   

  Yes, SpermFilter^® has been compared to another product on the marked (PureSperm®) in a comparative study of 898 ART cycles (245 IUI-H, 58 IUI-D and 595 IVF/ICSI cycles). There was no significant difference.
• What is the size of the silica particles in SpermFilter®?
  
   

  30-40 nm
• Does SpermFilter® contain antibiotics?
  
   

  No SpermFilter^® 100% stock solution does not contain antibiotics.
• What is the density?
  
   

  The density for 100% is 1.125 g/ml, 1.102 g/ml for 80% and 1.055 g/ml for 40%.
• Is SpermFilter® LAL tested?
  
   

  Yes. LAL (limulus amoboe lysate) is another word for an endotoxin test.
• What is the shelf life when SpermFilter® is kept under warm/hot conditions?
  
   

  When kept at 30-40 degrees C the shelf life is 1 year.
• What does v/v mean in connection with the gradients?
  
   

  % (v/v) means volume to volume in percentage. When making gradients from 100% SpermFilter^® it is important to keep the medium at room temperature. Example make 100 ml 80 % (v/v) SpermFilter gradient: Take 80 ml 100% SpermFilter^® and dilute it with 20 ml SpermWash^®.
• Why do the ready-to-use SpermFilter® gradients contain HSA when some of the other ready-to-use products do not contain HSA?
  
   

  We recommend using HSA in the ready-to-use gradients because research at ProVitro has shown, that you lose up to 25% of the motile spermatozoa when using gradient without HSA.
• Do SpermFilter® gradients contain antibiotics?
  
   

  Yes, SpermFilter^® gradients contain Gentamicin.
• What is the shelf life when SpermFilter® gradients are kept under warm/hot conditions?
  
   

  When kept at 30-40 degrees C the shelf life is 1 week for SpermFilter^® gradients.
• What is the shelf life for SpermWash® once the product has been opened?
  
   

  As long as you can work sterile and keep sterility of the product, the shelf life is 12 months from the production date. The expiry date is shown on the label.
• Can SpermWash® be recapped?
  
   

  The product should not be recapped. The Teflon-coated rubber stopper keeps sterility of the medium.
• How long before use should the bottle be removed from the refrigerator, if preheating is not available as an alternative?
  
   

  It depends on the size of the bottle. Larger bottles need more time than smaller. Generally it is advisable to remove the bottle from the refrigerator the day before use. The best option is to aspirate the volume needed, equilibrate it at the desired temperature. Place the remaining SpermWash^® in the refrigerator.
• If the bottle is not recapped how can it remain sterile ? Should we use a needle and syringe and puncture the membrane to get the SpermWash®?
  
   

  We advice you to use a needle and a syringe unless you are going to use the whole bottle at once. It is also advisable to wipe the membrane (rubber stopper) with a piece of cloth soaked in 70% ethanol. Wait for the ethanol to evaporate before puncturing the membrane with the needle.
• Why does the SpermWash® contain gentamicin?
  
   

  Gentamicin is active against both gram negative and gram positive bacteria as well as mycoplasma i.e. it has a broader spectrum than penicillin and streptomycin. Gentamicin is added to the SpermWash^® for the satisfaction of the customer. We cannot guarantee sterility, once the customer breaks the seal. . If the medium is contaminated, gentamicin will inhibit microbial growth.
• What does the SpermWash® kit contain?
  
   

  It contains 2 large and 1 small needles, two syringes and one sealed vial with SpermWash^® for preparing the swim up.
• What is the shelf life when SpermWash® is kept under warm/hot conditions?
  
   

  When kept at 30-40 degrees C, the shelf life is 1 week
• Can SpermCryo™ be used for freezing testicular tissues and semen that is oligospermic or should it only be used for freezing good quality semen samples?
  
   

  SpermCryo™ is only recommended for freezing high quality semen. For freezing patient semen, purified spermatozoa or testicular sperm we have developed the new SpermCryo™ All-round medium.
• What is the disadvantage of diluting the sperm too much?
  
   

  The volume increases so that you have to use more straws and you dilute the semen. When thawing, the volume can become too big to be able to load on one density gradient. It is especially important not to dilute the semen too much when working with a subnormal semen quality (teratozoo/oligozoo/astenozoo-spermia). It is generally not advisable to freeze semen below 1 mill/ml so a more concentrated extender (semen freezing medium) is better.
• After adding SpermCryo™ to the semen, how long is it necessary to leave the mixture at room temperature?
  
   

  For 10 minutes with occasionally mixing.
• How long is it necessary to leave the straws in nitrogen vapour?
  
   

  Our standard freezing procedure is as follows: We use a dewar approximately 25cm high. We place the straws horizontally in nitrogen vapour approximately 9 cm above the liquid nitrogen and 1 cm below the top of the dewar for 15 minutes at - 30°C. Then we lower the straws to a level approximately 6 cm above the liquid nitrogen for 15 minutes at - 70°C. After 15 minutes at - 70°C the straws are plunged into the liquid nitrogen.
• Can we wash the semen (SpermWash® + 5 min centrifugation -2500 rpm) before mixing it with SpermCryo™?
  
   

  When freezing purified and/or washed spermatozoa it is recommended to use our new SpermCryo™ All-round.
• Can SpermCryo™ be used on purified sperm samples?
  
   

  We cannot recommend using SpermCryo™ on purified sperm samples only on raw semen. Use our new SpermCryo™ All-round instead.
• Is it recommended to plunge the sample directly into liquid nitrogen?
  
   

  Direct plunging into liquid nitrogen is not an optimal choice. We recommend either a programmed freezing protocol or incubation in nitrogen vapour for 15-30 minutes before plunging into liquid nitrogen.
• What is the shelf life when the SpermCryo™ and SpermCryo™ All-round are kept under warm/hot conditions?
  
   

  When kept at 30-40 degrees C the shelf life is one year for SpermCryo™ and  one month for SpermCryo™ All-round.