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Expressed proteins and activated pathways in conditioned embryo culture media from IVF patients are diverse according to infertility factors.

ABSTRACT: OBJECTIVE:Given that the embryo culture medium secretome reflects the embryo development, we hypothesize that protein profiles are affected according to infertility factors, which can be responsible for detrimental embryonic developmental competence. The aim of this study was to screen the protein profile of conditioned embryo culture media in patients presenting deep infiltrating endometriosis (ENDO) and polycystic ovarian syndrome (PCOS) undergoing IVF, by proteomics approaches. The control group was constituted by tubal factor patients. METHODS:Patients underwent in vitro fertilization (IVF) treatment as routine and oocytes were fertilized by ICSI. The embryos were group cultured until day 3 of development, and after transfer the culture media were collected. For the proteomics analysis, two pools of samples were prepared for groups CONTROL and PCOS, and 4 pools of samples for group DIE. Samples were prepared to deplete high abundant proteins and followed evaluated by high throughput proteomics approach. RESULTS:The embryonic organ and tissue development were physiological functions activated, based on proteins identified in the 3 study groups of samples. The samples coming from DIE patients presented a high calcium activity and on the other hand, embryos coming from PCOS patients showed a decreased calcium action. Other pathways as grow factors through the EGF signaling pathway overexpressed in ENDO culture medium and protein kinase A in PCOS were also observed. CONCLUSIONS:Proteomic embryonic secretome will advance our knowledge of early embryogenesis and additionally could lead to improved selection of embryos for transfer warrants further investigation.

SUBMITTER: Bonetti TC

PROVIDER: S-EPMC6798601 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature

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Expressed proteins and activated pathways in conditioned embryo culture media from IVF patients are diverse according to infertility factors.

Bonetti Tatiana Cs TC Haddad Debora Cm DC Domingues Thais S TS Alegretti Jose Roberto JR Motta Eduardo LA E Seeley Kent K Silva Ismael Dcg ID

JBRA assisted reproduction 20191014 4

<h4>Objective</h4>Given that the embryo culture medium secretome reflects the embryo development, we hypothesize that protein profiles are affected according to infertility factors, which can be responsible for detrimental embryonic developmental competence. The aim of this study was to screen the protein profile of conditioned embryo culture media in patients presenting deep infiltrating endometriosis (ENDO) and polycystic ovarian syndrome (PCOS) undergoing IVF, by proteomics approaches. The co ...[more]

PMID: 31251011

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Project description:Study question: Which non-declared proteins (proteins not listed on the composition list of the product data sheet) are present in unconditioned commercial embryo culture media? Summary answer: A total of 110 non-declared proteins were identified in unconditioned media and between 6 and 8 of these were quantifiable. What is known already: There are no data in the literature on what non-declared proteins are present in un-conditioned (fresh media in which no embryos have been cultured) commercial embryo media. Study design, size, duration: The following eight commercial embryo culture media were included in this study: G-1 PLUS and G-2 PLUS G5 Series from Vitrolife, Sydney IVF Cleavage Medium and Sydney IVF Blastocyst Medium from Cook Medical and EmbryoAssist, BlastAssist, Sequential Cleav and Sequential Blast from ORIGIO. Two batches were analysed for each of the Sydney IVF media and one batch from each of the other media. All embryo culture media are supplemented by the manufacturers with purified human serum albumin (HSA 5 mg/ml). The purified HSA (HSA-solution from Vitrolife) and the recombinant human albumin supplement (G-MM from Vitrolife) were also analysed. Participants/materials, setting, methods: For protein quantification, media samples were in-solution digested with trypsin and analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). For in-depth protein identification, media were albumin depleted, dialyzed and concentrated before sodium dodecyl sulphate polyacrylamide gel electrophoresis. The gel was cut into 14 slices followed by in-gel trypsin digestion, and analysis by LC-MS/MS. Proteins were further investigated using gene ontology (GO) terms analysis. Main results and the role of chance: Using advanced mass spectrometry and high confidence criteria for accepting proteins (p< 0.01), a total of 110 proteins other than HSA were identified. The average serum albumin content was found to be 94% (92% - 97%) of total protein. Other individual proteins accounted for up to 4.7% of the total protein. Analysis of purified HSA strongly suggests that these non-declared proteins are introduced to the media when the albumin is added. GO analysis showed that many of these proteins have roles in defence pathways, for example 18 were associated with the innate immune response and 17 with inflammatory responses. Eight proteins have been reported previously as secreted embryo proteins. Limitations, reasons for caution: For six of the commercial embryo culture media only one batch was analyzed. However, this does not affect the overall conclusions. Wider implications of the findings: The results showed that the HSA added to IVF media contained many other proteins and that the amount varies from batch to batch. These variations in protein profiles are problematic when attempting to identify proteins derived from the embryos. Therefore, when studying the embryo secretome and analysing conditioned media with the aim of finding potential biomarkers that can distinguish normal and abnormal embryo development, it is important that the medium used in the experimental and control groups is from the same batch. Furthermore, the proteins present in un-conditioned media could potentially influence embryonic development, gestation age, birthweight and perhaps have subsequent effects on health of the offspring.

Dataset Information

Expressed proteins and activated pathways in conditioned embryo culture media from IVF patients are diverse according to infertility factors.

Publications

Expressed proteins and activated pathways in conditioned embryo culture media from IVF patients are diverse according to infertility factors.

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