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Inducing and culturing system-establishing method for totipotent stem cells quteASCs of mice or human beings

A technology of pluripotent stem cells and stem cells, applied in the field of induction and culture of totipotent stem cells quteASCs

Active Publication Date: 2017-05-31
INNER MONGOLIA UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] With the development of animal genetic regulation mechanism, gene editing technology, biological individual development and other life science basic theories and the development of bioengineering technology, animal stem cell induction methods have become more diversified and stem cell types have become more subdivided, but a " Totipotent stem cells” alone or through a chimera model can develop into an individual animal in the uterus like a normal fertilized embryo combined with a placenta

Method used

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  • Inducing and culturing system-establishing method for totipotent stem cells quteASCs of mice or human beings
  • Inducing and culturing system-establishing method for totipotent stem cells quteASCs of mice or human beings
  • Inducing and culturing system-establishing method for totipotent stem cells quteASCs of mice or human beings

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Embodiment 1

[0042] Embodiment 1: mouse novel totipotent stem cell (quteASCs) induction and establishment result

[0043] 1. Preparation of early mouse embryos

[0044]Obtain mouse embryos through natural mating and surgical methods, and recover 50 early embryos at 3.5 and 6.5 days after fertilization from the uterus of the treated mother mice respectively (in order to reasonably arrange the time of each experiment, the early embryos are usually recovered in the morning, calculated according to the time of fertilization Exactly 3.5 days and 6.5 days), used for new stem cell induction and culture establishment.

[0045] 2. ASCs transitional stem cell induction establishment

[0046] After the mouse blastocysts obtained by the above method for 3.5 days were cultured outside the KSOM culture liquid for 2 days, the zona pellucida was removed with Tyrode's acid and then inoculated into Fibronectin (fibronectin) (fibronectin was diluted to 1 with phosphate buffer solution). : 60 working soluti...

Embodiment 2

[0055] Embodiment 2: human novel totipotent stem cell (quteASCs) induction and establishment result

[0056] 1. Preparation of early human embryos

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Abstract

The invention discloses an inducing and culturing system-establishing method for totipotent stem cell quteASCs of mice or human beings. The materials are early embryo of mice growing for 3-8 days (obtained through in-vitro fertilization or collected from the uterus of a parent body), and the early embryo of the human beings growing for 3-14 days (obtained through in-vitro fertilization or collected from the uterus of a parent body). The inducing method comprises the following two key technical processes: 1, carrying out inducing to establish excess type stem cells ASCs, wherein the key inducing factor is AF; and 2, carrying out further inducing to complete the system establishing technological process of the novel pleuripotent stem cells quteASCs, wherein the key inducing factor is ABCL. According to the invention, through the specific inducing method, the novel stem cells quteASCs of the mice or the human beings are successfully obtained through the specific inducing method, and the totipotency of the novel stem cells quteASCs of the mice or the human beings for developing to the individual tissue of animal or the placenta of the animal is verified through the experimental means.

Description

technical field [0001] The invention belongs to animal and human reproductive and developmental bioengineering technology, and mainly relates to a method for inducing and culturing quteASCs of mouse or human totipotent stem cells. Background technique [0002] Animal embryonic stem cells (Embryonic Stem Cells, ES) research began in the 1990s, and was first successful in mice and humans. Two groups of scientists won the 2009 Nobel Prize for this. Animal adult cell stem cell induction technology (induced pluripotent stem cells, iPS) was founded by Takahashi and Yamanaka in 2006, using Oct4, Sox2, Klf4 and c-Myc four transcription factors to reprogram mouse fibroblasts to reverse development totipotent stem cells. In general, stem cell-related research carried out in various countries in recent years is mainly concentrated in the field of mice and humans. The mouse model is mainly for basic research on life sciences related to stem cells. The medical application of human stem ...

Claims

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Application Information

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IPC IPC(8): C12N5/0735C12N5/073
CPCC12N5/0696C12N2500/32C12N2500/44C12N2501/115C12N2501/155C12N2501/16C12N2501/235C12N2501/724C12N2501/998C12N2506/02
Inventor 包斯琴吴宝江李喜和
Owner INNER MONGOLIA UNIVERSITY
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