PureSperm 100 – 100 ml
For the preparation of density gradients
PureSperm 100 is a sterile colloidal silica suspension in an isotonic salt solution, optimized for the preparation of density gradients and used to separate and purify human sperm from semen for use in Assisted Reproduction Technologies (ART). PureSperm 100 effectively separates normal sperm from lymphocytes, epithelial cells, abnormal, immature and senescent sperm, cell debris, bacteria and seminal fluid.
- Increases sperm survival time.
- Removes sources of ROS.
- Removes bacteria.
- Removes sperm with abnormal DNA.
- Improves cryo-survival.
- Improves yield and recovery compared to swim-up techniques.
- Silane-coated silica
- Calcium chloride
- Potassium chloride
- Sodium chloride
- Purified water
- pH: 7.4-7.8.
- Osmolality (mOsm/kg H2O): 300-310.
- Endotoxin transfer during treatment: <1.0 EU/mL.
- Sperm survival 18 hours after density gradient separation: >70%.
Storage and Stability
- Store at 2 to 27ºC and avoid temperatures above or below these values. Under these conditions PureSperm 100 has a shelf-life of 24 months from production. The expiry date is shown on both bottles and cartons.
- Open and close bottles under aseptic conditions. After opening, store at 2 to 8ºC when not in use. Shelf-life on the product label applies when the product is stored according to manufacturer’s recommendations.
- No antibiotics, unstable additives or preservatives have been added by the manufacturer to PureSperm 100.
Precautions and warnings
- When retrieving the sperm pellet, follow the instructions given in the pack insert to avoid inadvertent contamination.
- Use aseptic procedures at all times.
- If available, use sealed buckets during centrifugation to avoid creation of aerosols.
- Clean accidental spills using a dampened cloth or paper. PureSperm 100 causes floors and benches to be extremely slippery.
- PureSperm 100 does not represent any kind of fire or combustion hazard. A material safety data sheet is available from the distributor or manufacturer.
- Do not use any solution which shows evidence of bacterial contamination.
- Do not use contents if tamper-evident seal is broken.
- Federal Law (USA) restricts this device to sale by or on the order of a physician.
- Please check for regulatory compliance governing the use of ART products in your country.
Prepare two PureSperm gradients for each semen sample. This reduces the risk of overloading a single gradient, provides security when handling tubes or recovering sperm pellets, and provides two tubes to balance the centrifuge rotor.
Comparative proteomic analysis of spermatozoa isolated by swim-up or density gradient centrifugation
Stefania Luppi1, Monica Martinelli1, Elisa Giacomini2, Elena Giolo1, Gabriella Zito2, Rodolfo C Garcia3† and Giuseppe Ricci12*†
Reproductive Biology and Endocrinology 2015, 13:36
Comparison of the DNA Fragmentation and the Sperm Parameters after Processing by the Density Gradient and the Swim up Methods
Iraj Amiri, Marzieh Ghorbani, Safora Heshmati
Journal of Clinical and Diagnostic Research. 2012 November, Vol-6(9): 1451-1453
Comparative study of the effects of three semen preparation media on semen analysis, DNA damage and protamine deficiency, and the correlation between DNA integrity and sperm parameters.
Charoenchai Chiamchanya, Nattpawit Kaewnoonual, Pachara Visutakul, Sirikul Manochantr and Jirattikan Chaiya.
Asian J Androl. 2010 Mar;12:271-277.
The use of two density gradient centrifugation techniques and the swim-up method to separate spermatozoa with chromatin and nuclear DNA anomalies.
Sakkas D, Manicardi GC, Tomlinson M, Mandrioli M, Bizzaro D, Bianchi PG, Bianchi
U.Hum Reprod. 2000 May;15(5):1112-6.
DNA fragmentation of spermatozoa and assisted reproduction technology.
Henkel R, Kierspel E, Hajimohammad M, Stalf T, Hoogendijk C, Mehnert C, Menkveld R, Schill WB, Kruger TF.
Reprod Biomed Online. 2003 Oct-Nov;7(4):477-84.
Recovery and survival of sperm is higher with PureSperm density gradient than swim-up in neat and cryopreserved-thawed semen specimen.
P Ranganathan, A Agarwal
Fertil Steril, 2001
Comparative study on density gradients and swim-up preparation techniques utilizing neat and cryopreserved spermatozoa.
Allamaneni SS, Agarwal A, Rama S, Ranganathan P, Sharma RK.
Asian J Androl. 2005 Mar;7(1):86-92.
Bacterial contamination and sperm recovery after semen preparation by density gradient centrifugation using silane-coated silica particles at different g forces
C.M. Nicholson L. Abramsson, S.E. Holm and E. Bjurulf
Human Reproduction, Vol. 15, No. 3, 662-666, March 2000
The use of PureSperm, mini-Percoll and swim-up preparation techniques to separate spermatozoa with chromatin and nuclear DNA anomalies.
M. Tomlinson et al
Abstract Andrology in the nineties 1999
Higher rates of recovery with PureSperm density gradient compared to ISolate
Agarwal, A. and Ranganathan, P.ESHRE 2001
A comparison of two density gradient preparations for sperm separation for medically assisted conception: Effect on recovery, clean-up, motility, and motion parameter.
J. A. Roudebush, et al.
Supplement of the 27th Annual Meeting of American Society of Andrology, 81 Seattle, USA (March/April 2002)
Comparison of Six Density Gradient Media for Selection of Cryopreserved Donor Spermatozoa
Nathalie Mousset-Siméon, Nathalie Rives, Lydie Masse, Florence Chevallier and Bertrand Mace
Journal of Andrology, Vol. 25, No. 6, November/December 2004
What should I use for diluting PureSperm 100?
PureSperm Buffer is the best media to use for dilution of PureSperm100. It has been optimised and ionically balanced to match PureSperm.
Is there any human serum albumin in PureSperm 100?
Research in our laboratory showed that the yield of motile sperm recovered from the gradient was approximately the same, regardless of whether or not the gradient layers contained human serum albumin. Therefore, we have not included 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, 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, therefore, will not 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?
Yes, you can use PureSperm Wash to dilute PureSperm. 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.
Do we need to incubate PS100 and PS40/80 in 6% CO², at 37°C before use?
The most important to avoid is temperature fluctuations for the sperm. The ejaculate is approximately around room temperature or slightly higher. Therefore, we recommend equilibrating PureSperm to room temperature, not more. The preparation is also done at room temperature and therefore the temperature is still maintained.