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SAGE media

By: A.B., 15 June 2011

we needed some opinion in working with a SAGE media. Our clinic performed 10 IVF/ICSI procedures and 40 IUI procedures using SAGE media,no one of patients became pregnant. Like as protein suplement was used HSA : for HTF,Q1 and Q2 5% and for HTF(HEPES) and ET 10%. Culture of embryos under oil in the drops. CO2 in incubator 6,0%. We discovered that the embryo development became slower, only one patients embryos was growing till blastocyst and form them only on day 6. Could be the problem related whith media delivery? Last order we receved media in warm condition and we decided not to use them,the previous orders was better,but I can't to name media "cold". Any opinions,please?

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Responses (6)
Response from chris 27 November 2011
If you are certain that there is no other laboratory factors, the most common of which is deviation from established procedures by technical staff, then the first thing you should investigate is your oil. Did you perform a bioassay such as sperm survival, not just an exposure study? I mean use a large dish with the oil in question and culture in microdrops proven fertile sperm and check its survival over 72 hrs. If sperm does well then measure pH of your culture media in the same conditions (microdrop or large volume, oil overlay or open) you would culture your human embryos. This would prove that the gas tank, the incubator and the media work corectly to give you the optimal pH. If you could not uncover anything after these two steps, it is time to clean your incubator and your lab, throw away the old lot of dishes and start new. Good luck!
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Response from Dr. krishna 20 June 2011
SAGE media yields optimal culture results with 5% co2. 6% is a lot and the pH significantly changes and that could be the reason for which the embryos are getting arrested at cleavage stage. Though SAGE comes with phenol red, its hard to pick pH changes and minor pH changes does matter in embryo culture. 6% co2 is more suited for COOk media.
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Response from 20 June 2011
have been using sage for quite a while with rather good results. check ph of media and then sdjust the co2 of the incubator to give the required ph. i have my incubator at 6.2%co2to give me a ph of 7.225 - 7.23
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Response from David Hickman 19 June 2011
We use Sage at a CO2 of around 5.0 - 5.2% in order to reach a pH of around 7.27-7.30. My suggestion is to check pH rather than CO2 concentration if at all possible. We have found that with different medias we need to adjust CO2 to reach the optimum target pH. When we first started using Sage, a CO2 of 6% put our pH too low for optimum development. Check the package insert to see what the target pH is for the appropriate media and adjust CO2 according to whatever works in your lab.
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Response from biju p 19 June 2011
Even I had same problem..SAGE medium is bad now a days,,,
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Response from ÷zhan 15 June 2011
Hi. I was used sage media( I hadn't any problem), but now baceuse of CE obligatin in my country I changed media. There isn't any wrong aplication you done. I think your problem is related with media delivery. I suggest that to you refresh your media.Second, always you have to another option for these circumstances. (you need back up media)Most important thing is, did you check incubator CO2 level?
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