Specification Of Primordial Germ Cells In Vitro And Identification Of Spermatogonial Stem Cells In Monkey Testis

Embryonic stem cells are highly undifferentiated cells.Inducing pluripotent stem cells refers to the reprogramming of cells into pluripotent stem cells with the characteristics and functions of embryonic stem cells.In mammals,genetic material is transmitted to offspring through germ cells,thus allowing the species to be continued.Embryos already form two types of cells in the early stages of development: somatic cells and germ cells.The first germ cells to be formed are primordial germ cells,precursor cells for sperm and eggs.In male animals,primordial germ cells gradually differentiate into spermatogonial stem cells as individuals develop.Spermatogonial stem cells eventually form sperms.This experiment is mainly divided into three parts:1.Mouse induced pluripotent stem cells were cultured in vitro and differentiated into mouse primordial germ cells.First,mouse induced pluripotent stem cells were cultured in a feeder layer system.When the cell state was stable,they were transferred to a feeder-free system.Immunofluorescence analysis of the induced pluripotent stem cells were mainly detected: OCT4,SOX2,SSEA4 and TRA-1-81.After the cell state was stabilized,they were induced into mouse epiblast stem cells,and then the mouse epiblast stem cells were induced into primordial germ cells.Finally,they were identified by immunofluorscence analysis,mainly detecting: OCT4 and Blimp1.2.Differentiate monkey embryonic stem cells into primordial germ cells.First,the monkey's embryonic stem cells were cultured in vitro.Immunofluorescence analysis of the pluripotency of monkey embryonic stem cells were mainly detected: OCT4,SOX2,SSEA4 and TRA-1-81.When the cell state was stable,they were induced into monkey na(?)ve embryonic stem cells.Immunofluorescence analysis of pluripotency of monkey na(?)ve induced pluripotent stem cells were mainly detected: OCT4,SOX2,TRA-1-60 and TRA-1-81.After the monkey na(?)ve embryonic stem cells were stable,they were induced into primordial germ cells.Finally,they were identified by immunofluorscence analysis,mainly detecting: OCT4,Blimp1 and SOX17.3.Identification of monkey spermatogonial stem cells: The monkey's testis were removed and paraffin-embedded blocks were made.The testis embedded block is made into slices.Finally,they were identified by immunofluorescence analysis,mainly detecting: GFR?1,Thy1,and VASA.Mouse induced pluripotent stem cells were induced into mouse epiblast stem cells,and then the mouse epiblast stem cells were induced into primordial germ cells.Monkey embryonic stem cells were induced into monkey na(?)ve embryonic stem cells,and monkey na(?)ve embryonic stem cells were induced into primordial germ cells.Spermatogonial Stem Cells were identified by GFR?1,Thy1,and VASA in Macaca fascicularis testicular.