In vitro maturation method and in vitro maturation culture solution for mouse naked ovum
An in vitro maturation culture and in vitro maturation technology, applied in the direction of germ cells, vertebrate cells, animal cells, etc., can solve the problems that naked eggs cannot be matured, discarded, and oocyte resources are wasted
Inactive Publication Date: 2014-06-11
ANHUI AGRICULTURAL UNIVERSITY
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Problems solved by technology
At present, because oocytes wrapped in cumulus cells can obtain high development quality and maturity rate when matured in vitro, naked eggs cannot be matured and are often discarded, thus causing a waste of oocyte resources
Method used
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Experimental program
Comparison scheme
Effect test
Embodiment Construction
[0019] 1. Experimental method
[0020] 1. Isolation and culture of naked mouse eggs
[0021] Naked egg washing medium: α-MEM + 10% FBS
[0022] Nude egg in vitro maturation medium: α-MEM + 10% FBS + 10ng / ml EGF + 1.5U / ml hCG + 1 mM / ml Glu + 0.23 mM / ml sodium pyruvate + 10 uM Forskolin
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Abstract
The invention discloses an in vitro maturation method and an in vitro maturation culture solution for a mouse naked ovum. The in vitro maturation method comprises the following steps: firstly, carrying out isolated culture on the mouse naked ovum; secondly, detecting the maturation of the mouse naked ovum; and thirdly, detecting the development capability of the matured mouse naked ovum. The in vitro maturation culture solution of the mouse naked ovum consists of alpha-MEM, 10 percent FBS (Fibroblast Serum), 10 ng / mlEGF (Epidermal Growth Factor), 1.5 U / mlhCG, 1 mM / mlGlu, 0.23 mM / ml sodium pyruvate and 10 muM Forskplin. According to the in vitro maturation method and the in vitro maturation culture solution for the mouse naked ovum disclosed by the invention, an in vitro high-efficiency mature development system of the mouse naked ovum is established by adding a chemical reagent correlated with oocyte in vitro maturation in a mouse oocyte culture system; according to the system, in vitro maturation of the mouse naked ovum can be effectively and manually regulated and controlled; in addition, the matured naked ovum can have the development capability of normal oocyte for forming embryon, thereby providing important data for the mouse to be used as an experimental animal model in the aspects of new variety culture, transgenic breeding, variety improvement and the like and providing the foundation for development of a human assisted reproductive technique in depth and breadth.
Description
technical field [0001] The invention relates to a mouse naked egg in vitro maturation method and a mouse naked egg in vitro maturation culture solution. Background technique [0002] Oocyte, also known as ovum, is the premise and foundation of a new life ontogeny. With the development of modern life sciences, oocytes are important materials for the cultivation of new varieties, transgenic breeding and variety improvement. For human reproductive health, the in vitro maturation and culture of oocytes is the basis for the birth of test-tube babies. Therefore, the in vitro culture technology of oocytes is the core technology of modern reproductive development. Oocytes exist on the ovary in the body, and the oocytes present on the ovary of postnatal animals stagnate in the germinal vesicle (GV) stage, no longer proliferate, and the number is limited. With the increase of age, the number continues to decrease. and eventually dry up. Therefore, unlike the male germ cells, sperm...
Claims
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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/075
Inventor 杨盼曹鸿国章孝荣张飞张运海刘亚李运生陶勇方富贵
Owner ANHUI AGRICULTURAL UNIVERSITY
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