The Stability Of Genetics Before And After Differentiation Of Human Embryonic Stem Cells Into Endometrium-like Cells

Endometrium of women in the child-bearing age is affected by estrogen andprogesterone. Its proliferation stag, production stage appear alternately duringmenstrual cycle. Endometrium rapidly going decidualization after embryoimplantation is one of the most important thing during women’s pregnency. Theappropriate endometrial thickness is necessary conditions for embryo implantation.Embryonic stem cells are rooted in blastocysts’inner cell mass which are highlyundifferentiated. They have the ability to proliferation infinitely and developmentpluripotent. Since their pluripotency,many scientists and doctors mentioned them andplace high hopes on them to make contribution to the regenerative medicine. It wasthought to play an important role in medicine. However, congenital，pathological,artificial factors damage the balance of endometrium，resulting thin endometriumwhich decreases the rate of embryo implantation. Women ended with infertility. Thiskind of situation is mostly encountered in assisted reproductive technology.Human embryonic stem cells can differentiate into any types of cellstheoretically including the germ cells. On account of this thought we believe that we could simulate an in vivo environment during the culture of human embryonic stemcells to differentiate them into endometrium-like cells. Till now,some of theacademics try to differentiate the somatic stem cells into endometrium-like cells andsucceeded. Some others rebuild the damaged endometrium of mice through embryostem cell transplantation. These studies are focused on the treatment or renovation ofembryo stem cells and somatic stem cells for thin endometrium syndrome.Meanwhile, our lab spend a long time on differentiating human stem cells intoendometrium-like cells in these two years. Our study involves in the differentiationprotocols and how some factors which plays roles in differentiation works toinvestigate a more efficient differentiation way.Gene microarray is a new technology to detect the Single NucleotidePolymorphism and can do the variation analysis between them. Through thistechnology we can clearly see the small change in DNA samples althrough analysisbetween the DNA before and after differentiation.Finally we could find out thefunction of the DNA which mostly changed based on several gene database. As far aswe know, there are now rarely some study on the stability of genetics before and afterdifferentiation of human embryonic stem cells into endometrium-like cells.Objective:Using a new method like SNP chips scanning to evaluate the stability ofgenetics before and after differentiation of human embryonic stem cells intoendometrium-like cells.Further more, to explore whether its safe to differentiateendometrium-like cells by this protocol and to refer some reference and statisticalsupport for the cells transplantation study.Methods:1.To culture the human stem cells till embryoid body stage then induce theminto endometrium-like cell through adding different dense of differentiation agents.Observe their cellular structure and morphology under microscope and immunofluorescence staining the cells to detect them under confocal scanning lasermicroscope whether they express keratin which is a specific protein of endometrium.2.Extract the DNA sample after differentiation. Due to their different regionsdivided the samples into4groups.These4groups have6samples. Using the SNPchips to scan them individually.Results:In this study,4groups6samples are included. They all meet the commitmentof the SNP chips detecting requirement. After the quality test,each sample is scannedand the datas in4groups are compared one by one. Group1and group2are the earlygeneration hESCs.After differentiation the number of diversity sites is2300. Whilegroup3represent for the late generation ones,the result of it is3770. And group4is agroup which is for a dynamic observation on the hESCs’ induction from an earlygeneration to the late.And the data of the4th group becomes6956.Conclusion:1.The application of adding17-beta estradiol and other differentiation factorto induce endometrial epithelial-like cells through embryoid bodies way could besuccessfully.2.Comparing the late generation cells with the early ones we find that the DNAsite mutation becomes more and more along with cell generation growing.3.It’s about7000mutation loci in20generations of the differentiation ofhuman embryonic stem cells into endometrial epithelial-like cells. The mutations ofexons located in chromosome1,7are the most significant.