Method for constructing pregnancy induced abortion drug screening model by using induced pluripotent stem cells and application
A technology for pluripotent stem cells and drug screening, applied in the field of biomedicine, can solve the problems of loss of microenvironment, destruction of tissue and organ structure, unfavorable embryo toxicology analysis, etc.
Pending Publication Date: 2021-02-26
河南省生殖健康科学技术研究院
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Problems solved by technology
However, it loses the microenvironment of tissue and cell development, destroys the structure of tissues and organs, and is not conducive to the toxicological analysis of the overall embryonic development
Method used
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Embodiment
[0034] 1. Culture of Embryoid Bodies
[0035] 1) Source of iPS cells: The source cells of this study were purchased from the iPS cell line of the Chinese Academy of Sciences (UE017C1), and the exogenous factor combination OCT4, SOX2 and KLF2 were introduced into normal human urinary epithelial cells by electroporation to reprogram to obtain iPS cells. After testing various stemness indicators, it meets the standards of human pluripotent stem cells.
[0036] 2) Three-dimensional suspension culture of iPS cells into EBs:
[0037] Transfer the cells obtained in the above steps to serum-free STEMdiff with Matrigel matrix instead of feeder cells TM In the culture medium, replace the iPS cell culture medium every day. After culturing for 4 to 5 days, when the iPS cell colonies grow to 70% to 80% in each well, use 0.5mM EDTA digestion solution (brand: Life technologies, 15575020) to directly digest the cells. After incubating at 37°C for 5-8 minutes, stop the digestion;
[0038] T...
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The invention discloses a method for constructing a pregnancy induced abortion drug screening model by using induced pluripotent stem cells. The method comprises the following steps: culturing the induced pluripotent stem cells in a three-dimensional suspension culture manner to form an embryoid body; and applying a drug to be screened to a culture medium of the embryoid body for exposure, and screening and / or evaluating the drug by analyzing the development index of the embryoid body in the culture medium containing the drug to be screened. The invention also discloses application of the method in pregnancy induced abortion drug screening. The model constructed by the method is based on development of embryoid body organs, the early pregnancy abortion process of the whole embryo is dynamically stimulated, the development morphology and the change of embryonic differentiation are observed, the change of the diameter of the embryoid body, the change of apoptosis in the embryoid body andthe development level of the embryonic layer of the embryoid body are detected, and the drug abortion toxicity evaluation index is determined according to the above standard in order to provide a reference for subsequent drug screening.
Description
technical field [0001] The invention relates to the field of biomedicine, and relates to a method and application of using induced pluripotent stem cells to construct a pregnancy-induced abortion drug screening model. Background technique [0002] With the opening of my country's "comprehensive two-child" policy, the number of elderly and high-risk women is increasing, and medication during pregnancy is often unavoidable. A systematic review of drugs during pregnancy shows that 60% to 90% of pregnant women need to use drugs, with an average of 2 to 4 drugs and a maximum of 8 drugs. About 79% of pregnant women have taken drugs with unknown effects on the fetus, which increases the probability of spontaneous abortion, premature birth and low birth weight infants. Therefore, effective screening of drugs with reproductive toxicity is crucial to achieve the goal of safe drug use in pregnant women. Traditional drug reproductive toxicity risk assessment is often based on the resu...
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IPC IPC(8): C12N5/073C12Q1/02
CPCC12N5/0603G01N33/5088G01N33/5026G01N33/5014C12N2506/45C12N2513/00C12N2503/02C12N2500/90C12N2533/90G01N2500/10G01N2800/368
Inventor 毋姗姗邓锦波赵良范文娟王艳丽位婷婷
Owner 河南省生殖健康科学技术研究院