Final Stem Cell

B?t ??u t? lec 7-13 Câu h?i ôn t?p stem cell: 1/ FGSCs 2/ diabetes cells (Beta cells) 3/ How to make ICSI & ROSI. Comperation of them 4/ Section of testis 5/ mô t? PGD and PGS 6/ Oogenesis From iPS and ES cells 7/ Product of transgenic animals 8/ Diff of AS and ES (adv and dis) 9/ scaffold of stem cell (polymerics of scaffold) 10/ 1/Female Germline Stem Cells: (Introduction, Isolation, Characterization, Maintenance, Demonstration, Application) (15pts) The Female Germline Stem Cells (FGSCs) were successfully isolated from porcine ovarian tissue by enzymatic digestion using collagenase and trysine and different adhesive selection since somatic cells are attached to the culture disk; meanwhile the FGSCs are floating in the culture media, then FGSC are cultured in vitro, in DMEMF/F-12 and N21free-serum supplemented. 2 populations of FGSCs, one with a cell diameter of 5-7 mircromet and the other with 10-12 micromet, were observed with spherical morphology and express specific germline characteristics (Vasa, stella, Oct4, c-kit). The cells become identical at 10-12 micromet after 10 days in culture, Oct4 expression was reduced in the cytoplasm and increased the size in the nucleus, forming groups of cells clustered around theca cells colonies. The large clumps of ovarian cells will be removed during cell passage by the filter of 40 micromet, and FGSCs were maintain for 1 month. After one month, following the gradual death of ovarian cells, FGSCs transfer onto mitocycin C treated MEF feeder layers for a long term culture with the addition of stem cells factors. Applications: Ch?ng minh in vivo Applications: Treatment female infertility : enabling the generation of new oocytes, especially for women with diminished ovarian reserves or premature ovarian failure. ( cho phép t?o ra các t? bào tr?ng m?i , ??c bi?t ??i v?i nh?ng ph? n? có l??ng d? tr? bu?ng tr?ng gi?m ho?c suy bu?ng tr?ng s?m . ) Fertility Preservation : can be isolated and preserved for women undergoing treatments like chemotherapy, which may damage their reproductive systems. • Differentiation to other cell types for cell replacement therapy. • Production of transgenic animals. • To understand molecular mechanisms of proliferation and differentiation of germ line stem cells. • Toxicity test and pharmaceutical purposes. 2/Stem Cells in the Treatment of Diabetes (Introduction, Using AS or ES to establish Stem Cells, Characterization, Maintenance, Demonstration, Application) (15pts) a potential approach to treating diabetes using human embryonic stem (ES) cells by directing their differentiation into insulin-producing pancreatic islet-like cells. The process begins with the isolation of ES cells from the ICM of a blastocyst, followed by their aggregation into embryoid bodies. These cells are then guided into Nestin-positive progenitor cells, which can differentiate into pancreatic lineages under N2 medium, bFGF, B27 media supplement conditions. In the pancreatic pathway, 1st discard bFGF and add nicotinamide results in the formation of insulin-secreting clusters, resembling functional islets. These differentiated cells have applications in diabetes treatment through transplantation to restore insulin production, as well as in drug testing and basic research to understand ?-cell biology. 3/ How to make and diff between ROSI and ICSI? (10pts) Application of IVF using mature sperm and immature sperm: If sperm motility is very weak, abnormal motility using ICSI - intracytoplasmic sperm injection. We are pick up sperm and directly inject it into the egg If the men no have sperm we need to collect sperm inside the testis, injected directly into oocytes and apply artificial activation by chemical or electronic is the ROSI method as a treatment for male infertility. Process ICSI and ROSI ROSI The difference between mature sperm and immature sperm: (5pts) x 5/ mô t? PGS and PGD PGD is an important option that should be offered to couples at risk of having a child with serious genetic disease. Genetic testing of an embryo for a specific gene mutation when one or both biological parents are al ready have the existence of genetic abnormality by examining a representative sample taken at a preimplantation stage of development. PGS: A preventative measure used to identify chromosomal abnormalities in the embryo, even if there’s no known evidence of a genetic abnormality in either parent. 6/ Oogenesis From iPS and ES cells, how to make this-lec 12 Reconstitution of female germ-cell development in vitro is a key challenge in reproductive biology and medicine. We show here that female (XX) embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) in mice are induced into primordial germ cell–like cells (PGCLCs), which, when aggregated with female gonadal somatic cells as reconstituted ovaries, undergo X-reactivation, imprint erasure, cyst formation, and exhibit meiotic potential. Upon transplantation under mouse ovarian bursa, PGCLCs in the reconstituted ovaries mature into germinal vesicle-stage oocytes, which then contribute to fertile offspring after in vitro maturation and fertilization. Our culture system serves as a robust foundation for the investigation of key properties of female germ cells, including the acquisition of totipotency, and for the reconstitution of whole female germ-cell development in vitro. Vi?c tái t?o s? phát tri?n c?a t? bào m?m cái trong ?ng nghi?m là m?t thách th?c quan tr?ng trong sinh h?c và y h?c sinh s?n. Chúng tôi trình bày ? ?ây r?ng các t? bào g?c phôi cái (XX) (ESC) và các t? bào g?c ?a n?ng c?m ?ng (iPSC) ? chu?t ???c c?m ?ng thành các t? bào gi?ng t? bào m?m nguyên th?y (PGCLC), khi k?t h?p v?i các t? bào sinh d?c cái d??i d?ng bu?ng tr?ng ???c tái t?o, s? tr?i qua quá trình tái ho?t hóa X, xóa d?u ?n, hình thành nang và th? hi?n ti?m n?ng gi?m phân. Khi c?y ghép d??i túi ho?t d?ch bu?ng tr?ng chu?t, các PGCLC trong bu?ng tr?ng ???c tái t?o s? tr??ng thành thành các tr?ng giai ?o?n túi m?m, sau ?ó góp ph?n t?o ra th? h? con cái có kh? n?ng sinh s?n sau khi tr??ng thành và th? tinh trong ?ng nghi?m. H? th?ng nuôi c?y c?a chúng tôi ?óng vai trò là n?n t?ng v?ng ch?c cho vi?c nghiên c?u các ??c tính chính c?a t? bào m?m cái, bao g?m vi?c ??t ???c tính toàn n?ng và ?? tái t?o toàn b? quá trình phát tri?n t? bào m?m cái trong ?ng nghi?m 7/Application of transgenic animal (summary) ATryn (Antithrombin Recombiant) for the prevention of blood clotting (?ông máu) in Antithrombin deficient patients Most common from genetically modified cows are lactoeferrin, breast milk produced from cows in the form of a supplement, and insulin. Insulin can also be made from genetically engineering cows. Insulin is more strong and beneficial this way. Insulin is used to treat diabetes EPO (Erythropoietin) is used to treat anemia or given to patients after chemotherapy. “ The most common use is in people with anemia associated with abnormal function (dysfunction) kidney. Many athletes, such as cyclists and runners, use EPO because the increasing number of red blood cells can increase oxygen capacity and produce improved physical activity 8/Diff of AS and ES (adv and dis) 9/ scaffold of stem cell (mô t? 2 hình) ? The AM is rich in Extracellular Matrix (ECM), displays numerous biological characteristics that render it highly desirable for biomedical applications. Because of its antiinflammatory and antimicrobial properties, the AM has been useful for its ability to reduce inflammation and scarring andenhance wound healing. ? Because of its low immunogenicity and tolerance by the host, the hAM has been used as a grafting material and bioscaffold in tissue regeneration, such as liver, bone, heart, and neurological repair. ? The hAM is rich in ECM and stem cells. Application for use as a bioscaffold necessitates decellularization to avoid immune reactions. Protocol for preparation includes alkaline decellularization of epithelial cells using NaOH. ? The hAM stem cell components of epithelial and MSCs make it also a rich source for stem cell extraction for purposes of regenerative medicine.