How long are frozen biopsied cells good?

[nuthinbutpink]

So, if an embryo is biopsied and the cell is frozen prior to testing, how long can the cell stay frozen before analysis? Embryos seem to do fine indefinitely but is it a different story with a single cell?

[Carole]

So, if an embryo is biopsied and the cell is frozen prior to testing, how long can the cell stay frozen before analysis? Embryos seem to do fine indefinitely but is it a different story with a single cell?

The cell is put in lysis buffer which dissolves the cell. The solution containing the dissolved cell can be frozen indefinitely so no worries on that score. Carole

[zibibbogirl]

Carole, I am wondering if all clinics use this lysis buffer method?

I am a little confused and I am worried I have given someone the wrong advice on the HT forum based on my experience with my clinic....

When I cycled in Australia I discussed batching in the event I did not have sufficient numbers to send to PGD. They advised me that if I wanted to batch they would biopsy the embryos from cycle one on day 5. They would then freeze the embryos and freeze the biopsied cells seperately and store them. I was told I would then need to cycle again within two months of the original cycle when the first batch of embryos had been biopsied. I was told that biopsied cells were not as stable as embryos and once biopsied had a limited timeframe in which they could be sent to PGD. I was told that if I was not prepared to cycle again so soon, I would be better off freezing the entire embryo intact from my first cycle, and then once I had completed my second cycle, thaw those out from the first cycle, biopsy everything together and then refreeze/freeze everything from both cycles and await PGD results.

My clinic sends out its PGD rather than doing it in house. I am wondering if the transportation of biopsied cells has anything to do with the method of freezing. Perhaps the dissolving method is not appropriate for transporting cells? Can you please clarify?

[Carole]

Carole, I am wondering if all clinics use this lysis buffer method?

I am a little confused and I am worried I have given someone the wrong advice on the HT forum based on my experience with my clinic....

When I cycled in Australia I discussed batching in the event I did not have sufficient numbers to send to PGD. They advised me that if I wanted to batch they would biopsy the embryos from cycle one on day 5. They would then freeze the embryos and freeze the biopsied cells seperately and store them. I was told I would then need to cycle again within two months of the original cycle when the first batch of embryos had been biopsied. I was told that biopsied cells were not as stable as embryos and once biopsied had a limited timeframe in which they could be sent to PGD. I was told that if I was not prepared to cycle again so soon, I would be better off freezing the entire embryo intact from my first cycle, and then once I had completed my second cycle, thaw those out from the first cycle, biopsy everything together and then refreeze/freeze everything from both cycles and await PGD results.

My clinic sends out its PGD rather than doing it in house. I am wondering if the transportation of biopsied cells has anything to do with the method of freezing. Perhaps the dissolving method is not appropriate for transporting cells? Can you please clarify?

Dear zbibbogirl,
I can only comment on my personal experience in research labs and clinical labs--which may not pertain to some of the newer techniques. Here's an article that talks about allele drop out and freezing before and after lysing the cell Detailed investigation of factors influencing amplification efficiency and allele drop (http://molehr.oxfordjournals.org/content/9/7/411.full), so there are differences in PGD protocols used over time and between labs. I would follow the advice of the program that you are working with. They are following the cell prep directions from the lab they are using. Good Luck. Carole