Spontaneous in vitro hatching of the human blastocyst: the proteomics of initially hatching cells
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Spontaneous in vitro hatching of the human blastocyst: the proteomics of initially hatching cells Miriam Almagor 1
&
Yishai Levin 2 & Rona Halevy Amiran 1 & Sheila Fieldust 1 & Yael Harir 1 & Yuval Or 1 & Zeev Shoham 1
Received: 24 August 2020 / Accepted: 19 October 2020 / Editor: Tetsuji Okamoto # The Society for In Vitro Biology 2020
Abstract Spontaneous in vitro hatching of human blastocysts starts with the formation of a tunnel through the zona pellucida (ZP) by cellular projections of trophoblast cells. Our aim was to identify the proteins that are upregulated in these initially hatching cells as compared to trophectoderm (TE) cells from blastocysts that had not yet hatched. Forty seven women that underwent assisted reproduction treatment donated their ICSI-derived polyploid blastocysts for the study. In polyploid blastocysts that started spontaneous hatching, hatched clusters of cells were collected from the outer side of the ZP. Liquid chromatography mass spectrometry was applied to determine the proteins that were upregulated in these cells as compared to TE cells obtained from inside the ZP. Whole non-hatched polyploid blastocysts were used as controls. Overall 1245 proteins were identified in all samples. Forty nine proteins were significantly upregulated in hatching cells and 17 in the TE cells. There was minimal overlap between hatching and TE samples; only serine protease inhibitors (SERPINS) and lipocalin were detected in both samples. Myosin and actin were highly upregulated in the hatching cells as well as paraoxonase, N-acetylmuramoyl alanine amidase, and SERPINS clade A and galectin. In the TE cells, gamma butyrobetaine dioxygenase, lupus La protein, sialidase, lysosomal Pro-X carboxypeptidase, phospholipase b, and SERPINS clade B and A were among the most highly upregulated proteins. These findings may contribute to the basic knowledge of the molecular behavior of the specific cells that actively perforate the glycoprotein matrix of the ZP. Keywords Human blastocysts . ICSI . Spontaneous hatching . Proteomics
Introduction Successful hatching of the human blastocyst is mandatory for implantation and establishment of pregnancy. In assisted reproduction laboratories, hatching of blastocysts on days 5–6 of culture is often observed (Hardarson et al. 2012). The activation of the hatching process is not dependent on endocrine
Miriam Almagor and Yishai Levin should be considered similar in author order * Miriam Almagor [email protected] 1
Infertility and IVF Unit, Kaplan Medical Center, affiliated with Hadassah Medical School, the Hebrew University, Jerusalem, Israel , POB 1, 76100 Rehovot, Israel
2
The Nancy and Stephen Grand Israel National Center for Personalized Medicine, Weizmann Institute of Science, Rehovot, Israel
effectors derived from the uterine milieu because it happens spontaneously in vitro. Usually spontaneous in vitro hatching is preceded by expansion of the blastocyst (Hardarson et al. 2012). This induces stretching and thinning of the ZP. Then, at one point, a single tro
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