Induction method of zebrafish induced pluripotent stem cells and inducible medium and ips medium used in the induction method

Abstract

The invention discloses an inducing method for danio-rerio inductivity multi-potential stem cells. The induction method includes the following steps that danio-rerio tissues or danio-rerio embryos are gotten, and danio-rerio fibroblasts are obtained through in-vitro isolated culturing; the danio-rerio fibroblasts are infected through viruses, the infected danio-rerio fibroblasts are induced, and the danio-rerio inductivity multi-potential stem cells are obtained through manual selecting cloning and subculturing. Based on the same technical scheme, the invention further discloses an inductive culture medium and an iPS culture medium used for the inducing method. RT-PCR analysis indicates that iPS cells induced through the embryo fibroblasts have the potency of being differentiated into various triploblastic-source cells. By means of the method, the inductive culture medium and the iPS culture medium, the technical difficulty that the infection efficiency of slow viruses in fish cells is low is overcome, and new tool cell sources and new concepts are provided for researching a danio-rerio somatic-cell reprogramming mechanism to select cell materials.

Description

technical field [0001] The invention belongs to the field of biotechnology, relates to the field of fish genetics and breeding, in particular to a method for inducing zebrafish induced pluripotent stem cells, an inducible medium and an iPS medium used in the method. Background technique [0002] Since Evans et al. and Martin established embryonic stem (embryonic stem, ES) cell lines from the inner cell mass (ICM) cells of mouse embryos in 1981, many researchers have tried to establish various mammalian and non-human cells. Mammalian ES cell lines. However, it took 17 years from the mouse ES cell line to the human (Homosapiens) ES cell line, and the human ES cell line was not established until 1998. Fish is the most diverse group of vertebrates, and researchers have also done a lot of research on its ES cells. The establishment of fish ES cell lines began in the 1990s, mainly focusing on medaka (Oryzias latipes) and zebrafish (Danio rerio), two small model species. In addi...

AI Technical Summary

Problems solved by technology

This is a very common iPS induction system, which has precedents in different species such as mice, pigs and humans. For example, Chinese patent CN 103923877 A provides a method for establishing a DOX-regulated porcine somatic cell induction reprogramming system , but currently this technology is limited to the induction of mammalian iPS cells, and there is no precedent for using this induction system to induce fish cells, especially fish cells with a higher degree of differentiation (lower developmental potential) (such as caudal fins)

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