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Preparation method of completely small molecule compound induced goat multipotential stem cells

A technology of small molecular compounds and pluripotent stem cells, applied in the field of stem cells, can solve the problems of biological safety risks and achieve the effects of short induction period, simple operation and clear chemical composition

Inactive Publication Date: 2018-02-23
GUANGXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, none of the above mediation methods can avoid the use of exogenous gene fragments, so all of them may bring biosafety risks to subsequent applications

Method used

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  • Preparation method of completely small molecule compound induced goat multipotential stem cells
  • Preparation method of completely small molecule compound induced goat multipotential stem cells
  • Preparation method of completely small molecule compound induced goat multipotential stem cells

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Embodiment 1

[0036] A method for preparing goat pluripotent stem cells induced by a complete small molecule compound, comprising the following steps:

[0037] 1. Isolation of goat ear margin fibroblasts and mouse fetal fibroblasts

[0038] Collect goat ear margin tissue pieces within 30 days from the goat farm, wash them three times with PBS buffer containing double antibodies, cut the tissue pieces with sterilized ophthalmic scissors, and place the small tissue pieces evenly on the culture dish In the process, turn it upside down and observe whether the tissue block is firmly attached to the wall in time, add an appropriate amount of high-glucose medium containing 10% FBS for culture, try to prevent the tissue block from floating, and then place it at 37°C, 5% CO 2 cultured in an incubator. The culture medium can be supplemented in an appropriate amount the next day, and then the medium is changed every two days. . Cells can be subcultured when they reach 90% confluence. Isolation met...

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Abstract

The invention provides a preparation method of completely small molecule compound induced goat multipotential stem cells. The preparation method comprises the following steps: separation of goat ear-rim fibroblasts and rat fetus fibroblasts, preparation of a feed layer, preparation of a cell culture medium, production of goat induced multipotential stem cell-like clone, recognition of morphological characteristics of goat induced multipotential stem cell-like clone and identification of biological characteristics. By the preparation method of the completely small molecule compound induced goatmultipotential stem cells, goat somatic cell reprogramming is induced under the condition of completely small molecule compounds and zero serum, the induced multipotential stem cells without virus vectors or exogenous gene integration can be obtained, and good cell materials are provided for follow-up research. The preparation method is simple to operate, facilitates repetition, and is short in induction period; ingredients of a compound are clear; and the safety is quite high. The form of the goat induced multipotential stem cell-like clone obtained by the preparation method is similar to that of rat ESCs.

Description

technical field [0001] The invention belongs to the technical field of stem cells, in particular to a method for preparing goat pluripotent stem cells induced by complete small molecule compounds. Background technique [0002] Embryonic stem cells (Embryo Stem Cells, ESCs) can be isolated and screened from the inner cell mass (Inner Cell Mass, ICM) of early embryos before implantation or fetal primordial germ cells (Primordial Germ Cells, PGCs). Ability to self-renew and differentiate into cells of the three germ layers. Mouse and human ESCs were obtained in 1981 and 1988, respectively. With the development of stem cell technology, other species including pigs, monkeys, rabbits, cats, rats and goats have also established ESCs. The only authoritative ESCs that are generally accepted are humans, monkeys, and a few rodent species. In terms of large livestock species, including goats, there is no recognized authoritative ESCs, which hinders the research and application of tra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/074C12N5/071C12N5/073
CPCC12N5/0696C12N2501/71C12N2501/999C12N2502/13C12N2506/1307
Inventor 黄奔李兰玉石德顺张丹丹郑贝贝任颜颜叶升刘树林
Owner GUANGXI UNIV
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