jagomart
digital resources
picture1_Cell Suspension Culture Slideshare 77474 | Education 11 24


 139x       Filetype PPT       File size 0.55 MB       Source: wd.vghtpe.gov.tw


File: Cell Suspension Culture Slideshare 77474 | Education 11 24
abstract embryonic stem es cells are typically derived from the inner cell mass of the preimplantation blastocyst and can both self renew and differentiate into all the cells and tissues ...

icon picture PPT Filetype Power Point PPT | Posted on 03 Sep 2022 | 3 years ago
Partial capture of text on file.
                                           ABSTRACT 
           •    Embryonic stem (ES) cells are typically derived from the inner cell mass of the 
                preimplantation blastocyst and can both self-renew and differentiate into all the 
                cells and tissues of the embryo. Because they are pluripotent, ES cells have been 
                used extensively to analyze gene function in development via gene targeting. The 
                embryonic stem cell is also an unsurpassed starting material to begin to understand 
                a critical, largely inaccessible period of development. If their differentiation could 
                be controlled, they would also be an important source of cells for transplantation to 
                replace cells lost through disease or injury or to replace missing hormones or genes. 
                Traditionally, ES cells have been differentiated in suspension culture as embryoid 
                bodies, named because of their similarity to the early postimplantation-staged 
                embryo. Unlike the pristine organization of the early embryo, differentiation in 
                embryoid bodies appears to be largely unpatterned, although multiple cell types 
                form. It has recently been possible to separate the desired cell types from 
                differentiating ES cells in embryoid bodies by using cell-type-restricted promoters 
                driving expression of either antibiotic resistance genes or fluorophores such as 
                EGFP. In combination with growth factor exposure, highly differentiated cell types 
                have successfully been derived from ES cells. Recent technological advances such as 
                RNA interference to knock down gene expression in ES cells are also producing 
                enriched populations of cells and elucidating gene function in early development. 
                                                                  
   FIG. 1. Development of the early mouse embryo from the E2.5 morula to the E6.5 gastrula...
                                                                                    Primitive endoderm
                                                                                      Epiblast
                               O'Shea, K. S. Biol Reprod 2004;71:1755-1765
                               Biology of Reproduction
                                                             
   Copyright ©2004 Society for the Study of Reproduction
            STEM CELL CHARACTERISTICS 
           •    The basic characteristics of a stem cell population: pluripotency, self-renewal. 
           •    transcription factors, maintain self-renewal and to inhibit differentiation, 
           •    STAT3, by the binding of leukemia inhibitory factor (LIF) to gp130 receptors. 
           •    Gene targeting experiments have demonstrated that neither STAT3, LIF, nor gp130 is 
                required to maintain the inner cell mass, although Stat3 –/– embryos die around E6.5, 
                and gp130 –/– embryos die after E12.5. 
           •    the essential POU homeodomain protein OCT4, the variant homeodomain containing 
                protein NANOG, the SRY family member Sox2, Foxd3 (previously Genesis) a member 
                of the forkhead winged-helix family, and possibly Wnt signaling. 
                                                                  
                                                       Oct-4 
           •    The Oct4 promoter contains a retinoic acid (RA) -responsive repressor element, such 
                that exposure of ES cells to the morphogen/teratogen retinoic acid inhibits Oct4 
                expression and allows expression of lineage-specific transcription factors present in ES 
                cells. The Oct4 promoter also contains a proximal element that drives Oct4 expression in 
                the epiblast that is downregulated at gastrulation, and a germ cell-specific distal 
                enhancer that is not active in the epiblast. Germ cell nuclear factor, an orphan nuclear 
                receptor, binds to the distal enhancer to restrict expression to the germ cell lineage. By 
                using this distal promoter to drive expression of EGFP to ES cells, then selecting those 
                cells, it has recently been possible to differentiate oocytes from mouse ES cells in vitro 
                [Hubner er al., 2003]. 
           •    OCT4 interacts with a number of other factors, including the SRY-related HMG box 
                containing transcription factor SOX2, E1A protein, the transcriptional coactivator Utf1, 
                and the Forkhead box protein FOXD3. 
           •    Several downstream targets of Oct4 have been identified, including the extracellular 
                matrix protein osteopontin, which may promote the lateral migration of primitive 
                endoderm to line the trophoblast as well as initiate expression of endoderm lineage-
                specific gene expression. Other targets include Hand1 and Fgf4, which are expressed in 
                early trophectoderm; Fbx15, which is expressed in ES cells and later in testis; and Zfp42, 
                also known as Rex1. 
                                                                  
                                                    NANOG 
            •    The recently identified pluripotency factor Nanog is a member of the homeobox family of 
                 DNA-binding transcription factors. Two human genes and a single mouse gene with three 
                 alternative splice variants have now been identified. Nanog is first expressed in the compacted 
                 morula, in the ICM, then in the proximal epiblast at the location of the future primitive streak. 
                 Nanog is downregulated at gastrulation in mesoderm and endoderm, remaining in the epiblast 
                 to E8. Nanog is present in ES cells, primordial germ, and EG cells and can be detected by 
                 reverse transcription-PCR in adult tissues as well.
            •    Interestingly, NANOG, STAT3, and OCT4 appear to affect both overlapping and independent 
                 events. 
            •    However, Nanog is expressed in Oct4 –/– embryos, suggesting that they act in parallel 
                 pathways. 
            •    Nanog null embryos and ES cells form extraembryonic endoderm at the expense of epiblast. 
                 Thus, like Oct4, Nanog appears to regulate both self-renewal and inhibit differentiation.
            •    Nanog appears to be critical at slightly later stages of development because deletion does not 
                 affect early stages of trophoblast differentiation (as is the case for Oct4) and it is expressed in 
                 the epiblast until E8, remaining in some adult tissues. Like Oct4, it appears that Nanog may 
                 regulate differentiation by transcriptional repression of genes that promote differentiation. In 
                 the case of Nanog, the enhancers of both Gata6 and Rex1/Zfp42 contain NANOG binding 
                 sites. Interestingly, NANOG contains an amino terminal region of homology to SMAD4, 
                 suggesting that NANOG in the epiblast may play a role in controlling TGFß signaling. 
                                                                     
The words contained in this file might help you see if this file matches what you are looking for:

...Abstract embryonic stem es cells are typically derived from the inner cell mass of preimplantation blastocyst and can both self renew differentiate into all tissues embryo because they pluripotent have been used extensively to analyze gene function in development via targeting is also an unsurpassed starting material begin understand a critical largely inaccessible period if their differentiation could be controlled would important source for transplantation replace lost through disease or injury missing hormones genes traditionally differentiated suspension culture as embryoid bodies named similarity early postimplantation staged unlike pristine organization appears unpatterned although multiple types form it has recently possible separate desired differentiating by using type restricted promoters driving expression either antibiotic resistance fluorophores such egfp combination with growth factor exposure highly successfully recent technological advances rna interference knock down p...

no reviews yet
Please Login to review.