The down-regulation of several genes involved in differentiation fat cell differentiation, heart and forebrain development, bone mineralization and segmentation are provocative and open the possibility that important alterations of fundamental process are taking place. In fact, knock out animals for several of these genes have very specific anomalous phenotypes. For example, Srf Arsenian et al. In addition, mutation in the Eomes gene, is shown to cause abnormal trophoblast development Russ et al.
In addition, Gata4 is able to induce visceral trophectoderm from ES cells Fujikura et al. Whether differences in gene expression will persist throughout embryonic development is not known. However, a group of genes involved in cardiac development Gata4 Kuo et al. It is possible that these changes are temporary and disappear, but could persist and affect long term development, extending the Barker hypothesis Barker to the preimplantation period. This indicates a specific effect on the TE cells either in terms of proliferation or survival in the IVF embryos.
Interestingly, a statistically higher number of IVF embryos developed from the two-cell stage to the blastocyst stage when compared with IVC embryos. These latter findings were surprising and contrary to our expectation. In fact, it is well known that embryos develop more slowly in culture when compared with the in vivo counterpart.
Additional time spent in culture, and in particular the first 24 h of development, would have been expected to slow the development even further. Our findings, however, suggest that a change in environment like the one present in IVC embryos fertilized in vivo and then cultured in vitro appear to be more stressful to the embryos; one could speculate that embryos benefit from being in the same environment from fertilization onward, no matter how artificial the initial environment is.
A change in environment represents a stressful shock, requiring time to adapt to the new condition. The timing of this particular stressor is important in determining its effect. It would be interesting to perform complementary experiments IVF and transfer of zygotes in the fallopian tubes to test this hypothesis.
Mutation of Nos1 has been shown to increase neurogenesis and mutation of Nrp1, Nrp2 down-regulated 1. Tgfb2, in addition to causing abnormal brain development, is associated with abnormal cardiovascular, respiratory, and reproductive morphology.
This indicates that although the IVF procedure shows fewer alterations in the level of specific gene expression when compared with IVC, some of the important morphogenic genes are altered in the IVF embryos. Overall, our new results confirm our past experiments.
The differences in gene number are likely due to the differences in laboratory environment, RNA amplification technique double amplification protocol versus linear amplification using the NuGen protocol used in the present experiments and the gene chip used in this study. In fact, in the prior experiments, we used the Affymetrix MOE A chip, containing only 22 transcripts.
The present set of experiments used the NuGen small sample amplification protocol because this protocol is robust and requires a reduced amount of starting RNA. In particular, the NuGen linear amplification protocol conserves the correct ratio between mRNAs that are expressed at different levels and does not distort transcript abundance.
Goff et al. Other authors have analyzed the effect of the fertilization method on mammalian embryos and their findings are complementary to ours. Whitworth et al. Interestingly, cluster analysis revealed that IVF and control embryos were different at the four-cell stage, but they did not separate in independent branches at the blastocyst stage, indicating a less dramatic effect of culture on embryos at this latter stage.
This could be a species- specific phenomenon. However, the two classes of embryos had significant differences: there were differentially expressed genes between the two groups and similar gene pathways were modified; for example, RNA metabolism and ribonucleoprotein complex were up-regulated in IVF embryos, a finding consistent with our observation.
Global pattern of gene expression in in vitro versus in vivo produced embryos was also analyzed by Corcoran et al. They found genes differentially regulated in IVF embryos. There was a general down-regulation of genes involved in transcription and translation, as well as genes involved in cellular metabolism.
One potential limitation of our experiments is the choice of WM, a medium that is known to be suboptimal. However, we specifically decided to use WM to stress the system and therefore, potentially magnify the effects of a different method of fertilization.
In this fashion, we wanted to glimpse the potential mechanism by which the method of fertilization affects embryo health. Future experiments will be conducted to assess the effect of an improved media and different methods of fertilization on the global pattern of gene expression.
Therefore, the results described could simply be explained by the fact that the blastocysts were at slightly different stage of development. In addition, the cell number differences only involve the TE cells, while the number of ICM cells is similar in all three groups.
If the blastocysts were truly at different stages of development we would expect the ICM number to be lower in the IVF group.
Indeed, we think that all the groups are in same stage of differentiation and the difference in cell numbers are secondary to increase incidence of apoptosis or decrease in cell proliferation, as confirmed by our microarray data. In conclusion, the in vitro produced mouse embryos in the specific culture conditions used are at a disadvantage when compared with in vivo embryos.
It remains to be investigated if these changes are only temporary or are associated with long-term consequences. For example, it is unclear if the down-regulation of the embryonic development genes is associated with delayed patterning in the cultured embryos. Presence of a long-term effect would be in agreement with the Barker hypothesis that holds that in utero events are associated with long lasting predisposition to disease Barker Another question to be elucidated is whether changes reported in one species might be present in another species.
More studies need to be performed to evaluate what is the effect of other forms of artificial fertilization i. ICSI on embryo development. Effect of fertilization method on blastocysts cell number and on the nuclear area of the ICM cells.
Gene ontology of family of genes statistically different between in vitro fertilization IVF and in vivo generated control blastocysts. Gene ontology of family of genes statistically different between in vitro fertilization IVF and in vitro culture IVC generated blastocysts. Real-time PCR verification of gene expression data. Expression profile of cyclin G1 Ccng1 , ubiquitin-conjugating enzyme E2A Ube2a and tousled-like kinase 2 Tlk2 genes in murine blastocysts produced by in vivo fertilization and culture in vivo , in vivo fertilization and in vitro culture IVC , and in vitro fertilization and culture IVF using both A microarray and B real-time PCR techniques.
Citation: Reproduction , 1; Number of transcripts differentially regulated according to the fertilization method. The number of genes either down- or up-regulated is the number that is statistically different after direct comparison of the described experimental group.
The exact number of genes is specified in the inferior portion of the graph. Hierarchical clustering and PCA analysis. A Hierarchical clustering analysis of all samples from different developmental stages or treatments. Unsupervised clustering in GeneSpring was used to analyze similarities among replicate samples across all treatment groups tested. Colors correspond to relative RNA abundance for the transcripts detected; each is represented by a horizontal bar in the heat-map.
Red indicates up-regulation, blue denotes down-regulation and yellow no change in expression. In vivo, IVF and IVC refer to blastocysts developed in vivo, cultured in vitro after fertilization in vitro and cultured in vitro after fertilization in vivo, respectively. B Principal component analysis of gene expression of all groups as in the hierarchical clustering analysis.
The authors declare that there is no conflict of interest that would prejudice the impartiality of this scientific work. The authors wished to thank Dr Emin Maltepe for his valuable suggestions. Human Reproduction 17 — Again, referring to what I said about overheating.
Now, sometimes after the transfer, there might be a little bit of blood from the instrument and on contact. So this is nothing worry about, don't panic. Now we often give you a pad for that purpose.
It doesn't mean a tampon is somehow bad for you, it just means sometimes a lot of times in fact, you're using vagina pessaries and topical medication. So it just makes things a bit messy, so we usually just give you a pad to use and the bleeding will never last for days on end. Okay, so finally, I think I need to address a very common question about acupuncture.
Now, there is no scientific proof acupuncture before or after transfer will improve your pregnancy right after IVF treatment but there's good evidence that acupuncture helps with mental wellbeing and certainly helps reduce stress level and anxiety levels associated with an IVF cycle.
So if you find that helps you by all means, go ahead. Because there's no proof that acupuncture is harmful to your IVF cycle. Okay, just want to recap, we talked a bit about what embryo transfer involves. And we also talked about how to look after yourself after the embryo transfer and we also talked about some of the myths that's surrounding embryo transfer. I certainly hope that you found this video helpful. Do remember, you all have your own individual circumstances and you should talk to your own doctor who know your full history.
Do leave us questions and comments and do subscribe to our channel so you can see other videos on fertility and fertility treatment. For now, it is goodbye until next time. It can be helpful to understand what happens on the day of your embryo transfer as the last step of an IVF cycle.
Find out more about IVF success rates Learn about other Fertility Treatments Understanding Infertility IVF is part of what we call ART Assisted Reproductive Technologies which allows fertility doctors to help people become pregnant if they cannot do so naturally. Read about the step by step process. The frustrations of infertility can place strain on mental health and relationships. Read our tips for self care here. Struggling to conceive is tough enough, but maintaining relationships while undergoing the ups and downs of fertility treatment has its own set of challenges.
What does the Medicare safety net mean? Video - How the IVF process works. Each IVF treatment cycle takes around six weeks. Step 1: Initial Specialist Appointment At your initial appointment, your fertility specialist will review your medical history and all previous investigations and treatments. Step 3: Treatment begins Your fertility nurse gives you the medication you need, explains the treatment cycle timeline, and shows you how to self-administer the Follicle Stimulating Hormone FSH injections.
Step 4: Hormone stimulation FSH is administered through a diabetic-style pen, stimulating your ovaries to produce more eggs than usual. Step 5: Treatment monitoring Throughout your cycle, regular blood tests measure your hormone levels and ultrasounds measure the size and number of your ovarian follicles. Step 6: Trigger injecton Once you have the optimum number and size of follicles, we plan your egg collection. Step 7: Egg collection in day surgery Egg collection is undertaken in day surgery, usually under ultrasound guidance.
Step 8: Egg fertilisation Collected eggs are taken to the laboratory and placed in culture medium to prepare them for fertilisation later that day. Step 9: Embryo development The egg and sperm are then placed in individual incubators at 37 degrees to mimic the temperature of the human body.
Step Embryo transfer Embryo transfer is a simple day surgery procedure and usually takes place five days after the egg collection. Step Embryo freezing Any extra embryos not used during a treatment cycle that are suitable for freezing can be stored for the future.
Step Pregnancy test Your nurse will organise an appointment for you to have a blood test two weeks after the embryo transfer. Antagonist treatment cycle Antagonist treatment uses injectable drugs called antagonists to prevent premature ovulation. The IVF protocols, explained. Welcome to Fertile Minds. How to prepare for embryo transfer day.
Want more information? IVF process infographic Download. Read the article. Managing relationships during IVF Struggling to conceive is tough enough, but maintaining relationships while undergoing the ups and downs of fertility treatment has its own set of challenges.
Request an appointment General enquiry. Request an appointment You must have JavaScript enabled to use this form. First name. Last name. Phone number. Email address. Shannon Hee Kyung Kim Prof. Michael Chapman. How did you hear about us? Subscribe to newsletter. If the patient does not respond to the first course, the dosage may be increased to mg; however, it is important to note that adverse effects are dose-related, and therefore the lowest possible dosage should be maintained.
Patients who do not respond after three courses should be reassessed. The onset of pharmacologic activity is seen in 5 to 10 days, and peak plasma concentrations are observed in about 6. Clomiphene is enterohepatically circulated, and its metabolites are excreted primarily in the feces via biliary elimination.
Although clomiphene limits the number of oocytes, it cannot be used alongside a GnRH analogue to avoid premature luteinization. Clomiphene should be used with caution in patients with polycystic ovary syndrome PCOS because of their increased sensitivity to normal doses of clomiphene and their risk for an exaggerated response. Pharmacists should caution patients who are taking clomiphene about the possibility of blurred vision and should advise them against driving if this occurs.
Patients should be reassured that symptoms of visual disturbance will resolve upon discontinuation of the drug. Patients may also experience hot flushes and ovarian enlargement. In the latter case, patients should be advised to avoid any activities that may cause trauma to the ovaries, such as pelvic examinations, sexual intercourse, and physical exercise.
These agents stimulate ovarian follicular production in women without primary ovarian failure and result in follicular growth and maturation. However, they have a narrow therapeutic window ranging from no effect to ovarian hyperstimulation syndrome OHSS. GnRH analogues should not be used in patients with primary ovarian failure that can be detected by high levels of FSH. GnRH analogues are also contraindicated in patients with overt thyroid or adrenal dysfunction, pituitary tumors, abnormal uterine bleeding of unknown origins, ovarian enlargement not caused by PCOS , or previous hypersensitivity to any of these agents.
Since their use leads to an increased risk of multiple pregnancies and spontaneous abortions, GnRH analogues should be used with caution and by experienced providers. The pharmacist should ascertain that the patient fully understands how to inject the medication correctly. Furthermore, to reduce injection-site irritation, the patient should be advised to rotate injection sites and to change needles after drawing up menotropins and before an injection.
If the patient experiences irritation at the site of injection, she should be advised to apply moist heat to the area. Other common adverse effects include abdominal cramps, abdominal swelling, abdominal pain, and headache. Since there have been reports of hypercoagulability in cerebral infarction related to OHSS, caution should be exercised when using GnRH analogues in patients who are predisposed to coagulation disorders. Repronex is produced via a step purification process, and the method for extracting Menopur is even more complex.
The initial dosage of Menopur is IU daily; after 5 days, adjustments of no more than IU per adjustment may be made. The maximum daily dose of Repronex should not exceed IU, and use beyond 12 days is not recommended. It is used in conjunction with hCG to produce ovulation in women who have previously undergone pituitary suppression. Adjustments to the dosage may be made once every 2 days and should not exceed 75 to IU per adjustment.
The maximum daily dose is IU per day. It is administered SC rather than IM and is as effective as follitropin in achieving ovulation induction and pregnancy. The code also includes rules which we have made known as directions to ensure procedures and practices are carried out properly, in line with the latest evidence.
We also include guidance on what high quality care looks like so that everyone can work towards achieving this. Read our Code of Practice. We want clinics and research centres to keep improving so we touch base with them regularly via e-newsletters and in person with the latest advice and information on how to provide the best quality care. No-one likes paperwork but fertility treatment is a big deal and should not be entered into lightly.
We produce these consent forms, and guidance for clinics on when and how to use them, to ensure consistency nationally.
View the consent forms and the associated guidance. How we regulate We regulate fertility clinics and projects involving research with human embryos.
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