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    Home > Biochemistry News > Biotechnology News > A simple, reliable and effective method of celoid cell enrichment.

    A simple, reliable and effective method of celoid cell enrichment.

    • Last Update: 2020-08-10
    • Source: Internet
    • Author: User
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    Recently, the Research Results of the Institute of Molecular Cell Science Excellence/Institute of Biochemistry and Cell Biology of the Shanghai Institute of Life Sciences of the Chinese Academy of Sciences, based on the title of Haploid Embryonic Stem Cells Be enriched and Maintained by Simple, was published online in Journal of Biological Chemistry.
    the study established a simple, reliable and effective method of meloblast enrichment.
    for more than a year, scientists have been working tirelessly on the exploration of monoploid cells.
    2011, mouse solitary female monoploid embryonic stem cells (parthenogenetic haploid embryonic stem cells, PG-haESCs) were established.
    2012, the mouse lone male monoploid embryonic stem stem cells, AG-haESCs, were established.
    followed, rats, monkeys, and humans were established.
    haploid cells because they contain only one set of chromosomes, do not cover recessive traits, can provide a great help in the study of genetic function;
    combine these traits, in the case of haESCs in mice, to bridge the gap not only in the cellular level but also at the individual level to study gene function.
    however, haESCs since birth is accompanied by the problem of constant spontaneous double-dip.
    spontaneous doubleple led scientists to try unsuccessfully to build haESCs from the very beginning of ESCs until streaming sorting techniques (fluorescence activated cell sorter, FACS) were applied to the enrichment of hapos.
    principle, after Hoechst staining, cells with different DNA content are distinguished under laser exposure and collected by electrocharged the monoploid cells to deflect them, thus enabling the enrichment of monoploid cells.
    but Hoechst dyes affect DNA replication to some extent.
    facS-enriched cells that grew far less than normal.
    the equipment and costs required for streaming, laser exposure throughout the process, power-up, and other pressures can bring additional potential genetic mutations to cells.
    , this method requires complex and expensive instrument support and is expensive.
    in this context, the Li Jinsong research team, based on the difference in the average diameter of the monoploid cells and the double-expanded cells, maintained the monoploid sonofability in vitro steadily by filtration method for more than 30 generations.
    the filtration system consists of a commercial membrane filled with 5 m or 8 m small holes and a support device for a membrane, plus a syringe.
    cost and can be aseptic.
    study of cell activity and single cloning formation has found that filtering first-generation cells is far superior to facS-rich cells.
    at the same time, filtering different numbers of AG-haESCs can still support the birth of semi-cloned mice.
    CRISPR/Cas9-mediated gene knockout and knock-in can be achieved normally on the filtered rich cells, and the corresponding gene knockout semi-cloned mice were born normally. in addition
    , the method can be used for the establishment of mouse haploid cells and the enrichment of human haploid cells.
    method of filtering emery cell-rich cells provides a guarantee for the development, application and popularization of monoploid cell-related technology.
    Yu Chao and Yu Meng are the co-first authors of the paper, and the research work has been supported by the Genome-wide Labeling Project (GTP), the Ministry of Science and Technology, the National Natural Science Foundation of China, the Chinese Academy of Sciences Strategic Pilot Science and Technology Special Project (B) and the Shanghai Science and Technology Commission;
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