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Culture method for development of waste embryo blastocysts

A technology for embryos and blastocysts, which is applied in the intersecting field of microfluidic technology and reproductive biology, can solve the problems of small number of ICM cells, difficulty in establishing lines, etc., and achieve the effect of improving the development rate of blastocysts

Pending Publication Date: 2020-06-02
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These abandoned embryos with stunted growth or poor morphology rarely develop to the blastocyst stage, and even if blastocysts are formed, the number of ICM cells is very small, which brings certain difficulties to line establishment

Method used

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  • Culture method for development of waste embryo blastocysts
  • Culture method for development of waste embryo blastocysts
  • Culture method for development of waste embryo blastocysts

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Preparation of Pluronic F-127 Modified Microfluidic Chip

[0043] 1) Soak the microfluidic chip in absolute ethanol for 15-40min, rinse with deionized water, dry in vacuum, and set aside;

[0044] 2) Put the cleaned microfluidic chip into the plasma generator, evacuate to 0-1Pa, introduce oxygen to 10-200Pa, adjust the discharge power at 5-300W, and perform radio frequency discharge for 0.5-10min;

[0045] 3) Apply a certain volume of Pluronic F-127 solution with a protein concentration of 0.2-2% on the surface of the chip in a vacuum state, and incubate for 2-6 hours (37°C) or 12-24 hours (4°C);

[0046] 4) Discard the chip surface solution in step (3), rinse 3 times with sterile deionized water, dry, and package.

Embodiment 2

[0048] Effect of Pluronic F-127 Modified Microfluidic Chip on Embryo Development

[0049] The microfluidic chip prepared in Example 1 and the microfluidic chip not modified by Pluronic F-127 were respectively soaked in 75% ethanol, placed in a vacuum incubator to vacuumize for 10 minutes, and then the embryo suspension was quickly mixed with 1×10 3 The density of embryos / mL is added to the surface of the chip, and a single embryo is introduced into a single trap by using negative pressure and gravity; when the embryo settles to the bottom of the trap, cover it with fresh medium and mineral oil to prevent evaporation; The medium is replaced every two days to supplement the embryos with nutrients and discharge metabolic waste.

[0050] The microfluidic chip modified by Pluronic F-127 above and the microfluidic chip of the control group unmodified by Pluronic F-127 were counted from the inoculation of embryos to start culturing, and the embryonic development including cleavage ra...

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Abstract

The invention provides a culture method for development of waste embryo blastocysts, and belongs to the cross field of microfluidic technology and reproductive biology. A chip used in the culture method is an ''inverted pyramid-shaped'' or ''inverted V-shaped'' chip with a concave array structure, and the surface of the chip is coated with Pluronic F-127. The embryo culture microfluidic chip is beneficial to realizing a single embryo positioning process and a culture process in vitro. According to the embryo culture chip and the culture method, the blastocyst development rate of waste embryoscan be obviously improved, and the embryo culture chip and the culture method have important significance for establishing human embryonic stem cell (hES) libraries of different human HLA types.

Description

technical field [0001] The invention belongs to the intersecting field of microfluidic technology and reproductive biology, and in particular relates to a culture method for the development of blastocysts of discarded embryos. Background technique [0002] Human embryonic stem cells (hES) are isolated and cultured from the inner cell mass (ICM) of blastocysts. They are cells with totipotent differentiation characteristics, self-renewal and multi-lineage differentiation potential. , Embryo development, drug screening, gene therapy and research on genetic and epigenetic mechanisms are of great significance. Human embryonic stem cell research began in 1998, and it has become the most active research field in life science after the Human Genome Project. In 1998, the Thomson laboratory of the University of Wisconsin in the United States used 36 fresh or frozen blastocysts donated by infertile couples for clinical treatment, and successfully isolated and established 5 hES cell li...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00C12N5/073
CPCC12M21/06C12M23/16C12M23/20C12N5/0604C12N2533/30
Inventor 秦建华苏文涛陈雯雯
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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