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A kind of measuring method of nuclear quantity

A measurement method and quantity measurement technology, applied in measurement devices, botanical equipment and methods, biochemical equipment and methods, etc., can solve the problems of cell development damage, uncertainty of enucleation amount, etc. Effects of damage to cell development

Active Publication Date: 2020-12-08
NANKAI UNIV
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  • Abstract
  • Description
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Problems solved by technology

[0003] The technical problem mainly solved by the present invention is to provide a method for measuring the amount of enucleation, which can solve the problem of the uncertainty of the enucleation amount in the blind aspiration enucleation operation and cause great damage to cell development

Method used

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  • A kind of measuring method of nuclear quantity
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Embodiment Construction

[0012] The cells used in this implementation example are domestic pig oocytes taken from a local slaughterhouse. The method of obtaining oocytes is as follows:

[0013] After the ovaries were removed from the slaughterhouse, they were transported to the laboratory within two hours in vacuum flasks filled with saline at 35° to 37°. Immediately wash twice with 37° sterile saline containing 100 IU / L penicillin and 50 mg / L streptomycin. The oocytes were extracted from the small sac with a diameter of 2 to 6 mm on the ovary, and the extracted cells were washed three times with TL-Hepes-PVA, and then matured in vitro (IVM) in an incubator at 39° with a carbon dioxide concentration of 5%42 Hour. After IVM, the cells were deovized with 0.1% hyaluronidase. Finally, the cells were washed three times with M199 to obtain the cells used.

[0014] The experimental operation of the present invention is based on the self-developed micro-operating system NK-MR601, and the camera is connect...

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Abstract

The invention discloses a measurement method of denucleation amount. The method is characterized in that cytoplasm in an entry needle can be automatically gathered to change the primary strip shape into an irregular ball shape after being released to an operation solution for a while during the blind absorbing denucleation process; the size of the cytoplasm ball can be measured to accurately obtain the denucleation amount. According to the method, the problem that the denucleation amount cannot be accurately estimated in the blind absorbing denucleation process is solved.

Description

technical field [0001] The invention relates to the technical field of somatic cell nuclear transfer. In particular, it relates to a method for measuring the amount of nucleation. Background technique [0002] Enucleation is to completely remove the main genetic material in the oocyte, so as to ensure that the individual developed from the transferred nucleus has the highest genetic characteristics similar to the nucleated animal (cloned animal). The amount of enucleation directly affects the outcome of later cell development. Incomplete enucleation can lead to aneuploidy of recombinant embryo chromosomes, resulting in polyploidy, abnormal cleavage, developmental arrest and early embryo death. If the enucleation is too much, it will accelerate the abnormal differentiation of the recombinant embryo and affect the embryonic development. Blind suction method is a commonly used enucleation method at present. The cytoplasm of the polar body and its lower 1 / 4-1 / 3 is aspirated n...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/873G01N33/483
CPCC12N15/873G01N33/4833
Inventor 赵新冯泽阳刘曜玮孙明竹崔茂盛
Owner NANKAI UNIV
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