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Method for establishing source of heredofamilial premature ovarian failure and inducing differentiation of totipotential stem cells to original genital ridge cells

A technology of totipotent stem cells and premature ovarian failure, applied in artificially induced pluripotent cells, biochemical equipment and methods, germ cells, etc., can solve problems such as failure to obtain oocytes

Active Publication Date: 2017-12-29
THE UNIV OF HONG KONG SHENZHEN HOSPITAL
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Problems solved by technology

Subsequent studies by many scholars have confirmed that human pluripotent stem cells can differentiate into oocytes or ovarian granulosa-like cells, but so far no oocytes with fertilization ability have been obtained.

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  • Method for establishing source of heredofamilial premature ovarian failure and inducing differentiation of totipotential stem cells to original genital ridge cells
  • Method for establishing source of heredofamilial premature ovarian failure and inducing differentiation of totipotential stem cells to original genital ridge cells
  • Method for establishing source of heredofamilial premature ovarian failure and inducing differentiation of totipotential stem cells to original genital ridge cells

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Embodiment Construction

[0029] The method for establishing familial premature ovarian failure-derived induced totipotent stem cells to induce differentiation into primordial germ ridge cells includes the following steps:

[0030] (1) Establish a familial premature ovarian failure disease model iPS cell line: Then use the combination of factors induced on mouse and human iPSCs, in 4i culture system (containing 3uM CHIR99021 (GSK-3 inhibitor), 1uM PD0325901 (MEK) Inhibitor), 5uM SB203580 (MAPK inhibitor) and 5uMSP600125 (JNK inhibitor) in the culture medium of 4 inhibitors) to ensure its totipotency before induction. Human iPSCs were infected with lentivirus carrying the VASA-GFP reporter gene (GFP inserted into the VASA promoter) for subsequent experiments.

[0031] (2) Induction and differentiation of human totipotent iPSCs into EpiLCs / PGCLCs and functional analysis of PGCLCs: In this protocol, we will refer to literature methods to culture in medium containing bFGF, TGF-β and 1% KSR for 2 days, and ...

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Abstract

The invention discloses a method for establishing a source of heredofamilial premature ovarian failure and inducing differentiation of totipotential stem cells to original genital ridge cells. The method is characterized in that adult cells of patients with heredofamilial premature ovarian failure are applied and reprogrammed into induced totipotential stem cells in vitro, and then are induced and differentiated into precursor cells of the oocytes, namely the original genital ridge cells. The method is a method for obtaining original genital ridge cells from patients with heredofamilial premature ovarian failure, lays foundation for further obtaining manure oocytes, and is expected to be applied in the field of reproductive medicine and translational medicine.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a method for inducing differentiation of induced totipotent stem cells derived from familial premature ovarian failure to primordial germ ridge cells, which lays a foundation for further obtaining oocytes. This method is expected to be applied in the fields of reproductive medicine and translational medicine. Background technique [0002] Currently, the incidence of infertility ranges from 3% to 17%. With the help of reproductive medicine doctors, only 70% of infertile patients get their own children. With the development of human assisted reproductive technology and the relaxation of the national two-child policy, more and more couples hope to have children with the help of assisted reproductive technology. However, clinically, some patients with premature ovarian failure and advanced age cannot obtain their own children due to the quality and quantity of oocyt...

Claims

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Application Information

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IPC IPC(8): C12N5/075G01N33/68
CPCC12N5/0609C12N2501/11C12N2501/115C12N2501/125C12N2501/15C12N2501/155C12N2501/235C12N2506/45G01N33/68
Inventor 杨胜彭淑平丁书芳徐千华熊琼帅词俊
Owner THE UNIV OF HONG KONG SHENZHEN HOSPITAL
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