Blastosis transfer

• Recently , there is a remarkable development of stage specific sequential culture media which allows the embryos to develop further in vitro for up to 5-6 days after egg recovery and that embryo stage so called "Blastocyst". The advantages of blastocyst culture are to eliminate those embryos with little developmental competence and to facilitate the synchronization of embryonic stage with uterine endometrial development. This means that we can expect the higher pregnancy and implantation rate from the blastocyst transfer. However , there is considerable patient variation , which means that on the same day using the same media , one patient may get 90% blastocyst development while another patient get only 10%. Additional research data found that about 5 % of patients having blastocyst culture have not at least one embryo develop to blastocyst so they end up with no transfer. 

Embryo Freezing

Embryo Freezing

If patients have many embryos, they can preserve their embryos for the next cycle by freezing. There are two methods to preserve embryos. The first method is slow freezing which will be used when patients want to freeze embryo on Day 1. The other method is Vitrification. This is the newest method of freezing embryos to reduce damages in embryos. Mostly we will use this method to freeze embryos on Day 5.

Why embryo freezing is important?

We recommend patients to freeze their embryos so that if this cycle of ICSI fails, they still have embryos enough to do Frozen Embryo Transfer (FET). The women do not have to go through all ICSI processes. Plus the price of FET is less than the whole ICSI program.

Sperm Freezing

Sperm Freezing Preservation 

Preservation
The sperm freezing preservation preserves sperm inside -196 °C in Liquid Nitrogen, which can be saved up to 10 years. For men that do radiation therapy, men that use birth control, or men that cannot give sperm collection to combine with the egg on the same day , the clinic is able to provide the preserve sperm with the sperm freezing.

Factors for a successful sperm freezing include:
•The sperm quality before freezing
•The sperm type and strength of protective layer
•Freezing and Soluble temperature

Surgical sperm retrieval

Surgical sperm retrieval is a treatment option for men with an absence or blockage of the tube (vas deferens) or non- obstructive azoospermia.

Sperm can be collected directly from the epididymis situated inside the scrotum (the pouch holding the testicles) where sperm are stored and mature using a fine needle and syringe. This is known as ‘percutaneous epidydimal sperm aspiration’ or PESA. Sperm can also be retrieved from the testicles, a process known as ‘testicular sperm extraction’.

Surgical sperm retrieval is carried out by a Consultant Uroandrologist who works in close association with the fertility unit and the procedure is timed to coincide with the female partner’s egg collection. If enough sperm is retrieved it may be possible to freeze small amounts for use at a later stage. The sperm collected is then used to inject the eggs using ICSI.

TESE: testicular sperm extraction. This involves opening up the scrotum and taking a large volume of testicular tissue, perhaps from several regions of the testicle. Sperm are then retrieved using a microscope to identify individual sperm.

TESA: testicular sperm aspiration. This involves placing a needle attached to a syringe through the skin of the scrotum and simply sucking out the fluid inside the testicle.

MESA: microsurgical epididymal sperm aspiration. An open surgical sperm retrieval procedure that uses an operating microscopy to locate the tubules of the epididymis precisely, so that large numbers of sperm can be extracted.

Sperm Preparation

• Sperm Preparation
  
  An ideal isolation technique should be rapid , inexpensive & isolate all the sperm without damaging them. IUI offers a simple and cost effective method of isolating the most functional spermatozoa from the ejaculate. It helps obtain an enriched fraction of motile spermatozoa and also  removes of seminal plasma and other cellular components from ejaculate. This is important because the cellular debris, prostaglandins and any micro organisms if present must be removed so that the sample can be used for IUI procedure. However, there is no technique, which is superior to all other techniques. Ideally, at the end of a sperm wash, we should aim to be able to attain minimum criteria of sperm quality in order to enhance the chances of pregnancy.
 
• 1. Simple Washing
• Washing is oldest method described. The aim is to remove only the seminal plasma, and not the cellular debris, bacteria or non-motile spermatozoa.
  After liquefaction, the semen is mixed with culture medium (1:1) and centrifuged at 300-400 g for 10 minutes. The supernatant is discarded and pellet is suspended in 2 ml of culture medium. This is centrifuged again at 300-400 g for 5-10 minutes and supernatant against discarded. The final pellet is suspended in 0.4-0.5 ml of medium and immediately used for insemination. This is especially useful when sperm density is very low and you are doing an IUI.
  2. Gradient Separation
• The layers are made with 1-2 ml of gradient solution (80% lower and 40% upper layer) 2 ml of semen is then carefully layered on top of 40% and centrifuged at 400-600 g for 15-20 min. cell debris, immobile an abnormal sperm all accumulate at interfaces and a soft pellet is formed at the bottom of the tube. This pellet is aspirated and suspended in 2 ml of culture medium (Depending on sperm density either simple washing or wash with swim-up technique is used). This is centrifuged at 300 g for 5-10 min. the pellet is now suspended in 0.3-0.5 ml of culture medium and used for insemination. The final wash with culture medium to remove the gradient is drawback of this method. Sedimentation Method / Layering under paraffin.
  3. Swim up
  • Swim – up form Pellet
  • This is the most widely used technique for separation of most sperms from non-motile sperms and cellular debris. It is used with normal semen samples and is based on principle active self-migration.
    After liquefaction, the ejaculate is mixed with culture medium and then centrifuged at 300-400 g for 10 minutes. The supernatant is discarded and the sperm pellet is gently over lay with 1 ml of culture medium and stored inside incubator at 37°C for 30-45 minutes. The 0.4-0.5 ml of the supernatant is then gently aspirated and used for insemination.
    Swim-up from Ejaculate
  • This is the ideal method because it obviates the needs for centrifugation . There are some workers who still prefer to centrifuge the recovered sperms.

Micro Array

Micro Array, PGS or a-CGH is another method of chromosome screening. However it is better than PGD as Micro Array can detect all 23 pairs of chromosome. The embryologist can see which chromosome is normal or abnormal. Due to Micro Array, we can screen and find the healthiest embryos for your family. Also we can know the gender of embryos. It works by detecting small amounts of missing or extra genetic material (DNA).Microarray is able to detect small copy number variants that were not detectable by previous methods such as PGD.

 

Assisted hatching

Assisted hatching

Another technique that our clinic provides to increase the chance of getting pregnant is Assisted Hatching. Before the embryos that are transfered back into the uterus can implant, embryos must hatch from the outer layer named Zona Pellucida. Our embryologist will make a little hole on the zona pellucida in order to make the hatching easier for the embryos.