Method of predicting endometrial receptivity

A technology of endometrium and receptivity, which is applied in the field of prediction of endometrial receptivity, and can solve problems such as inability to accurately determine specific participation

Pending Publication Date: 2022-06-07
HUDSON INST OF MEDICAL RES +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, ERA uses whole-tissue biopsies and therefore cannot precisely determine the specific involvement of specific cell types or specific molecules

Method used

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  • Method of predicting endometrial receptivity
  • Method of predicting endometrial receptivity
  • Method of predicting endometrial receptivity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0344] Example 1: Materials and Methods

[0345] Human endometrial tissue for isolation of primary endometrial epithelial cells

[0346] Ethical approval was obtained from the Human Ethics Committee at Monash Medical Centre (Melbourne, Australia) and all patients provided informed written consent. Endometrial biopsy samples were obtained from women who had undergone hysteroscopic dilation, curettage, or assessment of tubal patency. Menstrual cycle stages were determined by routine histological dates of tissues.

[0347] Isolation of Primary Human Endometrial Epithelial Cells (HEEC)

[0348] Tissues in the proliferative phase (days 6-14) were harvested into Dulbecco's modified Eagle's medium / F12 (DMEM / F12, Thermo Fisher Scientific, MA, USA) and cells were detached within 24 hours of collection. Cells were isolated by enzymatic digestion and filtration as previously described (Marwood et al., 2009). Briefly, collagenase (7.5 U / ml; Sigma) and DNase 1 (2000 U / ml; Roche, Castle...

Embodiment 2

[0411] Example 2: Proteomic identification of podocalyx proteins in primary human endometrial epithelial cells

[0412] As described in Example 1, primary endometrial epithelial cells (HEECs) from human endometrial tissue were isolated and enriched for plasma membrane proteins.

[0413] A total of 250 proteins were identified by mass spectrometry analysis of the obtained proteins (Table 2). Of these, 47 are considered to be cell membrane proteins, and 10 of them are involved in cell adhesion, including podocalyx protein (PCX).

[0414] To confirm the proteomic findings, total cell lysates of primary HEEC isolated from proliferating endometrium (for proteomic studies) were analyzed by Western blot using 3 antibodies directed against different regions of human PCX.

[0415] A dominant band of about 150 kDa was detected by all 3 antibodies, with a level of compatibility in both cell types. Ab1 detected an additional weaker band of about 80 kDa in HUVEC and HEEC, while Ab2 recog...

Embodiment 3

[0435] Example 3: PCX localizes to the apical membrane of human endometrial epithelial cells and endothelial cells and is specifically downregulated in the luminal epithelium, consistent with receptivity establishment

[0436] As described in Example 1, the cellular localization of PCX in the human endometrium throughout the menstrual cycle was examined by immunohistochemistry.

[0437] Similar staining patterns were detected for all 3 PCX antibodies. During the proliferative phase, PCX is strongly localized to the apical surface of luminal and glandular epithelial cells (LE and GE, respectively) and endothelial cells in blood vessels (BV). The matrix shows no / under detection. This pattern persisted more or less until early secretion, after which large differences emerged, especially in LE. In mid-secretory phase, PCX staining was barely detectable in LE, although still strong in both GE and BV. During late secretion, while LE continued to have minimal PCX, GE showed lighte...

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Abstract

The present invention relates to a method of predicting endometrial receptivity of an embryo implantation in a subject, the method comprising determining a level of podocalyxin in endometrial epithelial cells in the subject. The invention also relates to methods of monitoring epithelial cell receptivity and improving epithelial cell receptivity.

Description

[0001] Related application materials [0002] This application claims priority to Australian Patent Application No. 2019902204, filed June 25, 2019, entitled "Methods of predicting endometrial receptivity", the entire contents of which are hereby incorporated by reference. enter. [0003] sequence listing [0004] This application is filed with the Sequence Listing in electronic form. The entire contents of the Sequence Listing are hereby incorporated by reference. technical field [0005] The present disclosure relates to a method of predicting the endometrial receptivity of embryo implantation in a subject, the method comprising: determining the level of podocalyxin in endometrial epithelial cells of the subject. The present disclosure also provides methods of monitoring and increasing epithelial cell receptivity. Background technique [0006] Embryo implantation is a critical step in establishing a pregnancy, and failure to implant can lead to infertility. Assisted r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6876G01N33/68
CPCC12Q1/6876G01N33/689G01N2333/70596G01N2800/36C12Q2600/158C12Q2600/178C12Q1/6883G01N33/5091G01N2800/367
Inventor 聂桂英S·亨L·J·F·伦鲍茨
Owner HUDSON INST OF MEDICAL RES
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