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Method and quality control molecular based mouse embryo assay for use with in vitro fertilization technology

An embryo, assisted reproductive technology technology, applied in the field of quality control analysis of clinical assisted reproductive technology

Active Publication Date: 2016-03-23
FUJIFILM IRVINE SCI INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This striking transition is an initial, natural property of the early embryo and its homeostatic regulation during subsequent development in the later stages of preimplantation development that poses great challenges in the laboratory

Method used

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  • Method and quality control molecular based mouse embryo assay for use with in vitro fertilization technology
  • Method and quality control molecular based mouse embryo assay for use with in vitro fertilization technology
  • Method and quality control molecular based mouse embryo assay for use with in vitro fertilization technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0101] Example 1: Evaluation of regulators of pluripotency

[0102] Several factors such as the toxicity and sterility of the media or materials used in ART can affect the development of the embryo. The following examples describe assays for assessing embryo viability under optimal and suboptimal culture conditions. This example shows embryos in which regulators of pluripotency were visualized using fluorescence microscopy. Markers of pluripotency for evaluating embryonic development at the one-cell or two-cell stage of development include, for example, SOX2, Oct-4, Nanog, and their upstream mediators and downstream effectors, the upstream mediators and Downstream effectors play a role in ensuring normal embryonic development.

[0103] To test media, media supplements, or ART consumables, collected transgenic embryos were first incubated in 50 μL droplets of control media (the media identified to promote optimal blastocyst development) or test media. In each case, embryos w...

Embodiment 2

[0114] Example 2: Molecular Mouse Embryo Analysis

[0115] Transgenic mice containing reporter genes linked to pluripotency markers were used to perform molecular-based mouse embryo analysis (mMEA) under optimal and suboptimal conditions. Transgenic male mice expressing enhanced green fluorescent protein (EGFP) under the control of the POU protein domain class 5 transcription factor 1 (Pou5fl, also known as Oct-4) promoter and distal enhancer (B6; CBA -Tg(Pou5f1-EGFP)2Mnn / J; Jackson Laboratory).

[0116] Transgenic male mice were mated with superovulated wild-type (B6D2F1) female mice. Single-cell embryos were collected and cultured in groups of 3 to 4 embryos per 20 μL droplet of medium, or a single embryo per 10 μL droplet of medium, depending on the experiment performed. Embryos were cultured for 96 hours under optimal or suboptimal conditions. Fluorescence shows the expression of the Oct-4-EGFP transgene.

[0117] a. Comparison of morphological development and Oct-4 ex...

Embodiment 3

[0122] Example 3: Molecular Mouse Embryo Analysis for Evaluating Blastocyst Development in the Presence of Different Quality Oils

[0123] Transgenic mouse embryos containing CDX2-GFP were used to perform experiments in the presence of oils of different qualities ((1) good oil, (2) 5% low-quality oil, (3) 7.5% low-quality oil, (4) 10% low-quality oil, and (5) Molecular-based mouse embryo analysis (mMEA) under the condition of 15% inferior oil). Eighteen embryos were tested for each of the oil qualities listed above. Embryos were assessed morphologically for blastocyst development at 48 and 96 hours using a stereomicroscope, and also at 48 and 96 hours using a fluorescent microscope for GFP expression. Using conventional MEA procedures, any embryos (expanded and hatching blastocysts) with a 96-hour score of 80% or higher passed the test, and as such, the parameters of the test were considered achievable. As shown below, speciation analysis alone is not as sensitive as the mol...

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Abstract

A method for qualitatively assessing products used in in vitro fertilization is provided. Also disclosed is an improved quality control assay for use in clinical Assisted Reproductive Technologies (ART).

Description

[0001] Cross references to related patent applications [0002] This application claims priority to U.S. Provisional Patent Application No. 61 / 783,557, entitled "Methods and Quality Control for Molecular-Based Analysis of Mouse Embryos for In Vitro Fertilization Techniques," filed March 14, 2013, the U.S. The entire content of the provisional application is incorporated herein by reference. technical field [0003] The present invention relates to methods of evaluating products used in cell biology (eg, in vitro fertilization). The invention also discloses a quality control analysis method for clinical assisted reproductive technology (ART). Background technique [0004] In vitro fertilization (IVF) laboratories play an important role in treating infertile couples. Ensuring proper quality control (QC) in the IVF laboratory is critical to the success of any IVF program, as the laboratory environment can alter the quality of the embryos produced. Optimal media and a stable...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02A01K67/027G01N33/53
CPCA01K67/0275A01K2267/02A01K2267/0393C12Q1/68
Inventor 赫夏诺-滋·尼萨米拉·艾斯-萨拉米丽贝卡·苏珊·吉伯
Owner FUJIFILM IRVINE SCI INC
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