Research nad its pifalls

Pitfalls and Measures of Human Sperm Preparation Technique

We in this article like to explain all possible aspects related to pitfalls and measures of human sperm preparation technique. As well as some suggestion, how to  get best possible result. One by one our discussion will cover  –

  1. Pitfalls in field of research related to human sperm preparation technique.

  2. Factors affecting human sperm preparation technique.

  3. Pre – freeze semen parameters which decide the outcome.

  4. Other factors affecting human semen preparation technique.

  5. How to get best possible result.

Sperm preparation is broadly conducted  for two purpose.  Either by sperm processing or sperm freezing (cryo-preservation).  Both are having its own specific indication as well as method.  But having one  factor in common. That both method are useful and required to manage infertility. Sperm cryo-preservation (sperm freezing) is also used widely to preserve fertility in cancer patient. In this article “pitfalls and measures of human sperm preparation technique” our discussion will  describe social, scientific, future avenues of sperm preparation technique.   As well as  we will also cover other possible challenges  related to human sperm preparation technique .

Pitfalls in field of research related to human sperm preparation technique.

Like other field of  research,  sperm preparation technique is also facing lot of hurdles in the field  of research. First of all this field is not having  adequate research data and researchers even.  Low level of evidence is clearly  prevailing in this field.  Sufficient literature or documentation is not available . Secondly many literature are unavailable on the name of copy right, marketing policy or unaffordable cost.

Now a days in the field of human infertility  the greater part of the exploration centered around ovum and embryo. Or on the other hand as such research on sperm is ignored. While sperm play  equivalent role in infertility management . Just as sperms are similarly liable for prosperity of new generation.

Other very significant pitfall is unavailability of enough quantity semen sample for research work. Even in the developed country people are not coming forward generously to donate semen.  People are not discussing  seriously regarding semen or sperm preparation. Talking about semen or sperm may be a matter of fun.  But donation of semen is still not well accepted. Ethical and religious issues inversely affecting research process of sperm preparation.

Other pitfalls are legal issue.  Many country’s law do not allow  sperm processing and sperm banking. In some country payment for semen donation is legally not allowed.  Therefore in those places persons are not coming forward  for semen donation for research or therapeutic  purpose.

Also it is worth to mention here that presently researchers of this field are not in contact with each other.  Rather most of the manufacturer working in the field of sperm preparation are keeping their data reserve with themselves. Even wrong composition figure is being propagated to maintain secrecy and get better business.

Government owned organization and corporate working in the field of infertility do not like penetration of individual researchers in this field.  In other words  semen preparation  technique has been hacked for commercial gain. Every manufacturer and even government organizations  talk about their product and achievements with limited data.  But they do not provide clear figure.

One example  — Most of the manufacturer and  government organizations is performing MEA test with human sperm culture media. But nobody is giving any data about number of MEA test done with a particular batch.  Specially none of the manufacturing company talks about pitfalls of their product and technique.

Factors affecting human sperm preparation technique.

Proper training
How important is to have proper laboratory training about sperm preparation method.

The most important factor which  influence human  semen preparation  are   —-

  • Laboratory training.

  • Clinical disposables and media, like consumable plastic ware and compounds ( density gradient, semen extenders & cryo-preservative).

  • How and when to collect the semen sample ?

  • Each semen sample are different from other. It may in term of pH, sperm concentration, viscosity, debris, abnormal sperm etc. It is not practical to handle every semen sample by one technique or one compound.

Laboratory training of person handling semen for preparation is usually not up to mark. In usual course persons are taking embryologist training are handling semen for preparation. Average embryologist do not have proper training  of semen handling.  Rather presently focus of training is over ovum / embryo preparation.  It will not give a direct impact on  PR (pregnancy rate)

Proper cleaning and sterilization of plastic ware  is very much important. Usually  people are overlooking this aspect.  Taking good quality semen sample and using best possible compound (density gradient, semen extenders & cryo-preservative) is not all.

Collection of semen sample in morning hour gives a better result. Other factors  are  ejaculation abstinence, proper sleep, cleaning of genitals, humidity, etc.

Pre-freeze semen parameters which decide the outcome.

There are some pre-freeze factors of semen  which determine the outcome of human semen preparation.

Know about Pre-freeze parameter
How pre-freeze parameter decide outcome
  1. Quantity of semen sample

  2. Sperm concentration.

  3. Morphology of sperm — higher percentage of morphological abnormal sperm gives poor result.

  4. Viscosity & liquefaction time —  Highly viscous semen and  sample taking longer to liquefy gives poor result after preparation.

  5. Size of sperm — sperm with more cytoplasm, give unsatisfactory result after processing. Particularly after sperm freezing the outcome remain  poor in sperm with more cytoplasm. Excess of cytoplasm directly  affect motility. It can also produce excess harmful ROS.

  6. Dead sperm, debris  —  Dead sperm and debris produce ROS and affect all process and final outcome too.

    Other factors affecting human semen preparation technique.

Other deciding factors to affect outcome
Other factors which decide the result and PR
  1. General health of individual.

  2. Any chronic disease like diabetes, hypo-thyroidism, etc

  3. Intake of anti-cancer drug, steroid etc.

  4. Age of the person.

  5. Training of the person responsible for semen preparation and sperm freezing.

  6. Room temperature – Room temperature of 30 to 32 degree C gives best result.

  7. Temperature of incubator — For best result set the temperature of incubator between 32 to 34 degree C.  Semen is being ejaculated, deposited and transported for fertilization at 37 degree C. But its development , maturation and storage is at 3 to 4 degree C less than core body temperature.

  8. Humidity and temperature of environment– If both temperature and humidity is high, sperm quality and even the outcome will be poor.

  9. Incubation period of semen preparation technique — longer the incubation time  – poor the outcome.

  10. Plastic ware used for whole process.

  11. In case of cryo-preservation of sperm — rate of cooling and thawing process.

  12. For human sperm freezing, still we use slow freezing method which require costly equipment, programmable freezer. By developing an effective sperm vitrification method outcome will certainly  improve. As well as it will also make the process easy.

How to get best possible result.

For the future avenue of human semen processing technique, it is best to make co-ordination among researchers. As well as  save and make available the correct data. Hiding anything on behalf of copy right or marketing policy should be discouraged at all level.

Research on sperm require more attention. Because presently it is being an overlooked part of infertility management. May be on the ground that  work on ovum or embryo gives more business  to manufacturer.

Commercialization of research process is not having any way for better outcome.  And if possible it should be banned.

Persons should have desire to donate the semen. Legal factors affecting semen donation and any semen processing do not need any  consideration  on scientific and humanitarian ground.

Use of Human serum albumin (H.S.A.) is of no way safe in the human sperm preparation technique.  At some stage it will be a need and requirement to discontinue the use of H.S.A..  The main drawback with the use of H.S.A.  is transmission of disease causing viruses. Particularly emerging and un-diagnosed viruses.

Each semen sample are different from other. Therefore we should handle the sample according to its quality and other parameters.

For every human semen processing laboratory,  it is advisable to have their own semen  processing compound ( density gradient, semen extenders & cryo-preservative). Using blindly any manufactures product or data can never give a best or desired result.

Declaration of interest

The material contained in the article is original.  We are  not in process to publish or submit this article anywhere.  The author report no conflicts of  interest. The authors along with his organization “Frozen Cell: are  alone answerable for the material  and composing the article.

Acknowledgements  —  Frozen Cell Team is in a continuous process to create an best possible  environment against  ” Pitfalls and Measures of Human Sperm Preparation Technique”. The aim is to optimize  an ideal environment of research outcome  and process of human sperm.  We are in an undertaking to find a better strategy to preserve and process human sperm  by various procedure.

Funding  —  This post is solely prepared by Frozen Cell.  Any other organization or any system is not at all involved financially  in any way  for  this post.

Limitations —   We work with basic equipment and infrastructure.  Therefore not able to provide sophisticated  data.

Reference  :——

  1. Techniques used for IUI: is it time  for a change?  Human Reproduction, Vol.32, No.9 pp. 18351845, 2017.

  2. Process and Pitfalls of Sperm Cryopreservation, Section of Investigative Medicine, Imperial College London, Hammersmith Hospital.

  3. Keene, D.J.; Sajjad, Y.; Makin, G.; Cervellione, R.M. Sperm banking in the United Kingdom is feasible in  patients 13 years old or older with cancer.

  4. 4Wasilewski-Masker, K.; Seidel, K.D.; Leisenring, W.; Mertens, A.C.; Shnorhavorian, M.; Ritenour, C.W.;  Stovall, M.; Green, D.M.; Sklar, C.A.; Armstrong, G.T.; et al. Male infertility in long-term survivors of pediatric cancer  .


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