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APS TechNotes See all articles

Pure Water For In Vitro Fertilization

In Vitro Fertilization

APS Water Services has been heavily involved in the production of pure water for in vitro fertilization and cell culture applications at many prestigious centers.

The main use of pure water for cell culture is in the production of cell culture media. Media is essentially artificial blood that surrounds the cell and keeps it nourished until the  embryo is implanted back in the uterus. Since a petri dish and media do not contain an immune system or a blood purification system such as kidneys, any contaminant in the water used to make the media could enter the cell and cause it to die.  Media does contain an antibiotic that can take care of some bacteria, but over all the cell is not protected.

Contaminants to eliminate :

1. Dissolved ions

This would be basic ions like sodium, calcium and bicarbonate. If these ions are not removed then the Ion levels in the media would not be easily set. If you start with no ions in the water then you know exactly how much to add to the media.
These contaminants are removed with deionization.  

2. Dissolved organic

This includes anything in the water that was once alive and dissolved waste products or humans and animals. Frequently tap water contains high levels of organics and are toxic to cells.  Organics or removed using carbon, special ion exchange and specialty ultraviolet lights.

3. Bacteria

All tap water contains bacteria. Even if bacteria is removed it will grow back rapidly if not handled as a sterile product. Bacteria can give the embryo an infection and therefore it must be removed. Bacteria is removed using a 0.22 micron filter with a sterile tip.

4. Endotoxin/Pyrogens

Pyrogens also known as endotoxins are pieces of cell wall that left after a bacteria cell dies. Pyrogens cause hyper sensitivity reactions in Mammals and will certainly interfere with embryos.
Removal of pyrogens is essential and control of bacteria within the entire system will ensure low endotoxin levels in the final product water.  The main cause of pyrogens in water systems come form bacteria growth in storage tanks. Water should never be stored either before or after  it is purified because stagnant water will always grow bacteria unless it is in a sterile environment.  Bacterial growth will add Organic and Endotoxin contamination to the final product water.  If you are not getting optimal cell yield or your pregnancy success rate are not as high as you would like, you should remove any storage tanks on your water system.

Endotoxin is reduced using ultrafiltration. Ultrafiltration will not remove 100% of the endotoxin so it is important to keep the levels under control prior to an ultrafilter membrane.  

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