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Artificial Embryos
- 1. Presented By: Anjali Mandal Roll No.: BT17008 Course: Btech BT Sem-06 Submitted to: DR. Nilesh Wagh
- 3. Artificial embryos: • The artificial embryos were made by coaxing stem cells to spontaneously form tiny ball-shaped structures. • It include the beginnings of an amniotic sac and the inner cells of the embryo (the part that would become a person's limbs, head, and the rest of their body) though they lack tissues needed to make a placenta https://www.earth.com/news/embryo- structures-fetal-development/
- 4. Need Of The Hour: The discovery could pave the way to creating entire human embryos out of human skin cells, without the need for sperm or eggs These findings mean there may be no need to 'sacrifice' a live embryo to create a test tube embryo It could also have “vast implications” for modeling embryonic defects and shedding light on placental dysfunctions, as well as solving certain infertility problems by creating human embryos in a petri dish Researchers in future plan to generate embryos from sterile men and women, using only their skin cells, and generate a real embryo in a dish and implant the embryo in the mother
- 7. What are embryos? • An embryo is an early stage of development of a multicellular organism. • In general, in organisms that reproduce sexually, embryonic development refers to the portion of the life cycle. • It begins just after fertilization and continues through the formation of body structures, such as tissues and organs. A male human embryo, seven weeks old or nine weeks' gestational age https://en.wikipedia.org/wiki/ Embryo
- 10. Function: Epiblast The cell lineage that differentiates into 3 germ lines. Trophectoderm The cells that develop into the placenta. Extra embryonic endoderm cells The cells that develop into the yolk sac.
- 11. The challenge of creating embryos from iPSC The iPSC’s can be differentiated into the cells of all three germ layers- endoderm, mesoderm, ectoderm The main issue of recreating a complete embryo from iPSC is the requirement of other supporting cells- trophectoderm cells, extra embryonic endodermal cells Hence researchers had to find a way to first create these supporting cells types that helps the embryo to develop
- 12. What did they do? They used a novel approach to produce: i. Induced pluripotent stem cells (iPSC) ii. Induced trophoblast stem cells (iTSC) iii. Induced extra embryonic endoderm stem cells (iXEN)
- 13. How did they do it? https://www.sciencedirect.com/science/article/pii/S1934590919301171
- 14. The “GETMS” GATA-3: This gene is present in the embryo at the peri-implantation stage, 4- cell stage. Its found that in further developmental stages it is only found in trophectoderm cells. Eomes: Plays a crucial role during development. Functions in trophoblast differentiation and later in gastrulation, regulating both mesoderm delamination and endoderm specification. Tfap2c: It is known to be important for specification and maintenance of trophoblast stem cells (placental progenitors), but whether this factor also plays roles at later stages of placental development is less well understood.
- 15. The “GETMS” Myc: They are thought to function in the processes of cellular proliferation and differentiation Esrrb: It plays a vital role in placental development. Also, this factor is necessary for the maintenance of pluripotency in these cells. Moreover, the expression levels of “Esrrb” is found to be essential in deciding the fate of the cells (either into iTSC or iXEN)
- 16. Media used: MEFs: DMEM supplemented with 10% FBS, 2mM L-glutamine, and antibiotics. ESCs and iPSCs : DMEM supplemented with 10% FBS, 1%non- essential amino acids, 2mM L-glutamine, in-house mouse Leukaemia inhibitory factor, 0.1mM β-mercaptoethanol and antibiotics. Stable iTSCs: GFR-Matrigel-coated dishes in TX (tissue extract) medium. XEN cells and iXENs: RPMI supplemented with 15% FBS, 2mM L-glutamine, 1mM b-mercaptoethanol , in-house mouse FGF and 1 μg/ml heparin
- 17. Procedure: Take mouse embryonic fibroblast (MEF) Introduce TF- Gata3, Tfap2c, Eomes, Myc, Esrrb Viral transfection Nuclear Reprogramming iPSC iTSC iXEN
- 18. Study of fetus & placenta True embryonic stem cell iPSC’s injected into blastocysts to form an embryo using OKSM MEFs injected into blastocyst to form embryo using GETMS KEY: OKSM: Oct-4, Klf4, Sox2, Myc GETMS: Gata3, Eomes, Tfap2c, Myc, Esrrb https://www.sciencedirect.com/science/article/pii/S1934 590919301171
- 19. Drawbacks: To use this approach in human infertility treatments are: Proper 3-D scaffold to culture the cells is not yet available The right ratio between cell types and their environment are still unknown