Method for forming oocytes by induced differentiation of cattle induced pluripotent stem (iPS) cells

A technology of oocyte and cell induction, applied in germ cells, animal cells, vertebrate cells, etc., to achieve the effect of strong regenerative ability and plasticity

Inactive Publication Date: 2012-09-12
ANHUI AGRICULTURAL UNIVERSITY
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Problems solved by technology

Therefore, the directional differentiation of bovine iPS cells into oocytes has broad application prospects and development potential in cattle disease-resistant breeding, mammary gland bioreactor production, and protection and development of excellent genetic resources. There is no relevant report on the research of differentiation into oocytes

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  • Method for forming oocytes by induced differentiation of cattle induced pluripotent stem (iPS) cells
  • Method for forming oocytes by induced differentiation of cattle induced pluripotent stem (iPS) cells
  • Method for forming oocytes by induced differentiation of cattle induced pluripotent stem (iPS) cells

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Embodiment Construction

[0018] (1) Production of porcine follicular fluid

[0019] The pig ovaries used in the experiment all come from slaughterhouses. Since pig ovaries are gonadal tissues, pig ovary tissues after slaughter cannot be used for food, but they are used for scientific research on reproductive development, which is of great significance. The collected ovaries were placed in normal saline containing penicillin and streptomycin at 37°C and sent back to the laboratory within half an hour. Spray and disinfect the surface of the ovary with 75% alcohol, then wash it three times with 37 ℃ normal saline containing penicillin and streptomycin, and use a sterile syringe to extract the follicular fluid from follicles with a diameter of 6 mm on the surface of the ovary. The extracted follicular fluid is removed by centrifugation. After the tissue cell residue fragments were collected, the follicle supernatant was carefully aspirated and frozen for later use.

[0020] (2) Bovine iPS cell culture

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Abstract

The invention discloses a method for forming oocytes by induced differentiation of cattle induced pluripotent stem (iPS) cells, comprising three steps of pig follicular fluid preparation, cattle iPS cell culture and the directional differentiation of the oocytes of cattle iPS cells. In the method, the cattle iPS cells are differentiated into oocytes under definite induction condition. As only a small amount of cattle skin cells are inoculated to induce the cattle iPS cells, the application of the cattle iPS cells can effectively avoid the disadvantage that the separation of cattle embryonic stem (ES) cells needs to damage a large amount of excellent embryo. Thus, the method has wide application prospect and development potential in the aspects of cattle disease resistance breeding, mammary bioreactor production, protection and development of excellent genetic resources and the like.

Description

[0001] technical field [0002] The invention relates to a method for inducing differentiation of bovine iPS cells to form oocytes. Background technique [0003] iPS cells (induced pluripotent stem cells), also known as induced pluripotent stem cells, have the same developmental totipotency as ES cells (embryonic stem cells), and are of great significance in human regenerative medicine and livestock breeding and improvement. iPS cells have strong regenerative ability and plasticity in vitro, and can differentiate into female or male germ cells under certain induced environmental conditions. Therefore, in-depth research and application of iPS cell-related technologies can speed up the genetic breeding of livestock and the development of excellent breeds. protection and development. In addition, the production of transgenic animals is a hot spot in current life science research. Animal transgenics have important application prospects and development potential in biopharmaceu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/075C12N5/071
Inventor 曹鸿国章孝荣张运海陶勇刘亚李运生
Owner ANHUI AGRICULTURAL UNIVERSITY
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