The principal idea of this technique is to have more GM-CSF and other growth factors, but the critical thing is how to know the concentrations precisely to avoid any contamination, alteration or apoptosis? How can we evaluate that? Any suggestions?
It is probably better to avoid co-culture! You can obtain high blastocyst rates and blastocysts of good quality in semi-defined culture conditions (at least in the bovine species). To increase the quality of the blastocyst it is also better to avoid the use of serum (or to reduce it to 5% or less). The problem with co-culture is that the somatic cells and the embryos do not have the same needs at the same time. Moreover, it is much more difficult to control and it takes time to prepare the cells each time. At the UNCEIA (France) they used the Vero cells with good results. It is safer than primoculture at a sanitary point of view and easier to prepare. Hope this helps.
It seems ti be better to avoid co-culture, but Vero Cells could be the best choise in co-culture with blastocystes.
I think it depends on what species you mean. Human, bovine etc. The answer is not easy. However, in the human, autologous granulosa cells might be the best choice for safety. For best cells in co-culture in particular species the only way to answer is a good literature survey ..
I'm working on human embryos but in bad quality with hope to find a solution to make their qualities better until blastocyst stage so i thought about co-culture system with autologous cells (cumulus, lymphocyte)... But i still lost!!
Please see my paper, you will find the usefulness of using defined medium in getting high blastocyst ratio.
http://www.ncbi.nlm.nih.gov/pubmed/21220156
Before thinking about co-culture it can be useful to analyse the whole culture system adopted, e.g. culture media, but also gas purity, filters to lower TOCs, reduced oxigen tension, mini incubator, use of oil (this can be easily toxic). The blastocyst's rate is obviously conditioned by the oocyte and embryo developmental competence but this can be conditioned by any of the factors included in the culture system
May be co-culture is not the real solution. Perhaps there is a need to improve on the stimulation regime, handling during collection of oocytes, their handling in the lab, their fertilization method and subsequent handling of the resultant embryos. The culture medium used, the culture technique and so forth. My feeling is an improvement in some or all of the above can assist you obtain good quality embryos, blastocysts and acceptable pregnancy rates. In short the real problem thus could be total quality management. We now have commercial culture media that could provide you with quality embryos and blastcysts. So one really does not have to resort to co-culture technique that is fraught with a number of risks such as contamination of culture system, more man hours, possible transmission of disease especially when using donor cells instead of autologous cells, etc, etc. Improve the total quality management in your unit not just the lab but also the clinic. If you get bad oocytes due to poor stimulation or bad handling during oocyte collection you will not get good embryos even if your culture technique is good. I hope this is helpful. Good luck.
Jaffar Ali's answer has mentioned all the key points. Bad Oocytes = Bad Embryos. So improve the oocyte quality by improving the stimulation regime. If the poor quality is caused by poor genetics, there's nothing you can do to improve the base quality. Once you have the oocytes, ensure that everything they (and the resultant embryos) are subjected to in the laboratory is top quality: culture media, ALL culture conditions (including cuture vessels, mineral-parrafin oil, incubators, gas conditions, air quality in the lab), handling & manipulation. Extensive references (textbooks, peer-reviewed research) are available on these issues, especially for human.
Some Questions:
1. Why do you have 'bad quality' embryos? Are they excess embryos not used clinically that you are using for research, or are all your embryos of 'bad quality'?
2. With the co-culture research, are you trying to answer a specific biological question or are you simply trying to improve the quality of embryos that you use clinically?
Cell co-culture is not particulalry successful for embryo culture. AS someone else has stated, the somatic cells and embryos have different requirements, and of course the embryo's requirements change as they develop. However, if you were to use co-culture for a specific research project with key questions in mind, it's hard to better biology. So cumulus cells for fertilisation, oviductal cells for cleavage stages and endometrial cells for late morula-blastocyst.
i'm working in my project of thesis about in vitro maturation of oocyte specially for OPK women and i observed during my research that all OPK women even with mature oocyte MII after stimulation, those are bad and as consequence they have all embryo 'bad quality' so the important thing is not the quantity but the quality which is so low for those patients ...
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