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Differences in the expression of minor histocompatibility (Hm) alloantigens on two mouse embryonal carcinoma (EC) cell lines and the PYS-2 and T.D.M.-1 differentiated derivatives have been demonstrated by their ability to elicit a cytolytic T-lymphocyte (CTL) response. Experiments involving the use of various responder-target strain combinations on the one hand and Recombinant Inbred (RI) mice strains on the other have shown that: (i) there are major differences in Hm expression on the EC cells compared with the differentiated derivatives whose Hm expression appears more akin to that of adult splenocytes; (ii) although both EC cell lines show reduced Hm immunogenicity compared with adult splenocytes, major differences in the expression and possibly presentation between the F9 and PCC3 EC cell lines can be detected both by in vivo priming and by in vitro cold competition target experiments. These results are discussed in connection with the unexpected finding that some EC cell lines are capable of specific competition effects for appropriate CTL effectors despite their inability to stimulate such effectors in vitro and the absence of major histocompatibility complex (MHC) products. 相似文献
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O. F. Gordeeva 《Russian Journal of Developmental Biology》2014,45(4):224-230
TGFβ family factors play an important role in regulating the balance of self-renewal and differentiation of mouse and human pluripotent stem and embryonic teratocarcinoma cells. The expression patterns of TGFβ family signaling ligands and functional roles of these signaling pathways differ significantly in mouse and human embryonic stem cells, but the activity and functional role of these factors in mouse and human embryonic teratocarcinoma cells were not sufficiently investigated. Comparative quantitative real-time PCR analysis of the expression of TGFβ family factors in mouse embryonic stem, embryonic germ, and embryonic teratocarcinoma cells showed that embryonic teratocarcinoma cells express lower ActivinA than pluripotent stem cells but similar levels of factors Nodal, Lefty1, TGFβ1, BMP4, and GDF3. In human nullipotent embryonic teratocarcinoma PA-1 cells, most factors of the TGFβ family (ACTIVINA, NODAL, LEFTY1, BMP4, and GDF3) are expressed at lower levels than in human embryonic stem cells. Thus, in mouse and human nullipotent teratocarcinoma cells, the expression of ActivinA is significantly reduced compared with embryonic stem cells. Presumably, these differences may be associated with changes in the functional activity of the respective signaling pathways and deregulation of proliferative and antiproliferative mechanisms in embryonic teratocarcinoma cells. 相似文献
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We have induced teratocarcinomas from female embryos heterozygous for electrophoretic variants of the X-linked gene coding for phosphoglycerate kinase (PGK). An embryonal carcinoma cell line, P10, has been isolated from such a teratocarcinoma. It has a normal female karyotype and cultures contain both PGK isoenzymic forms. Clonal populations derived from P10 also contain both PGK electrophoretic variants. In addition, both X chromosomes in these cells replicate in synchrony with the autosomes during early S phase of the cell cycle. These data indicate that the undifferentiated P10 embryonal carcinoma cells contain two active X chromosomes. When cultured under the appropriate conditions, the P10 cells differentiate to form a variety of tissue types. At least some of these differentiated cells contain an inactive X chromosome as determined by cytogenetic analysis. Apparently X chromosome inactivation accompanies the differentiation of these female embryonal carcinoma cells. 相似文献
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Three protein products of the Duchenne muscular dystrophy (DMD) gene were identified so far. These include the two very similar muscle and brain type dystrophins, which are encoded by 14-kilobase (kb) mRNAs, and Dp71, which is much smaller. Dp71 is encoded by a 6.5-kb mRNA, which is transcribed from approximately 6% of the giant dystrophin gene. The present investigation shows that Dp71 is the first product of the DMD gene detectable during development. It is already expressed in the pluripotent embryonic stem cells. The two 14-kb mRNAs encoding the dystrophins are detectable only after differentiation of specialized cell types. The possible implication of these findings with regard to the ontogenetic activation and the evolution of the DMD gene are discussed. 相似文献
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Altered hormonal responses: markers for embryonal and embryonic carcinoma stem cells and their differentiated derivatives 总被引:1,自引:0,他引:1
Teratocarcinoma cells in culture offer an in vitro system for studying certain aspects of embryonic differentiation. To gain some insight into regulatory systems that might be operative during early development, we have characterized the alterations that occur in the hormonal responsiveness of the membrane adenylate cyclase of different embryonal carcinoma cell lines with differentiation. Each undifferentiated embryonal carcinoma stem cell studied (F9, PCC4, PC13, P19) has an adenylate cyclase system predominantly activated by calcitonin. Of great interest is the fact that cAMP production is also enhanced specifically by calcitonin in an embryo-derived stem cell line. Differentiation of the embryonal carcinoma stem cell population toward parietal endoderm results in a decrease in calcitonin activation with a concomitant appearance of sensitivity to parathyroid hormone. Differentiation toward visceral endoderm is characterized by a lack of response of the adenylate cyclase system to both calcitonin and parathyroid hormone. These results indicate that the changes noted in adenylate cyclase hormonal responsiveness might serve as useful markers during early stages of differentiation. 相似文献
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Protein patterns of developmentally totipotent mouse teratocarcinoma cells and normal early embryo cells 总被引:5,自引:0,他引:5
Protein patterns of mouse teratocarcinoma stem cells were compared, by two-dimensional gel electrophoresis, with those of early embryo cells. These malignant cells were known from previous experiments (B. Mintz and K. Illmensee, 1975, Proc. Nat. Acad. Sci. USA72, 3585–3589) to be capable of conversion to normalcy and of contributing to embryogenesis when introduced into a blastocyst. The protein comparisons were intended to reveal whether totipotent teratocarcinoma cells most nearly resemble normal totipotent cells of a specific stage, as a possible clue to their developmental origins. A simple method was devised for the purpose of generally facilitating comparisons of two-dimensional gels, among which technical variations commonly alter the absolute positions of individual proteins. This variation was normalized by the use of a reference constellation, or a network of lines connecting shared landmark proteins identified in all the gels. Whereas the network may undergo topological change from one gel to another, it continues to provide a readily recognized standard of reference. Protein patterns displayed many similarities and some differences, hence nonidentity, between teratocarcinoma cells and all normal preimplantation embryo stages tested, as well as between the various embryo stages themselves. The results also unexpectedly disclosed, however, that changed physiological states or posttranslational alterations may contribute significantly to some of the protein differences irrespective of the developmental status or potentialities of the cells. For example, in the OTT 6050 teratocarcinoma transplant line, pure teratocarcinoma cell groups (“cores”) found in the ascites fluid synthesized several proteins not expressed when the cores were enveloped (in embryoid bodies) by a yolk saclike epithelium; yet the core cells from both sources form comparable tumors if injected subcutaneously and are able to undergo differentiation if injected into blastocysts. In another comparison, some proteins that were present in inner cell masses isolated from blastocysts were absent in intact blastocysts, possibly because of their modification by the surrounding trophoblast in the latter case. These observations imply that protein differences between embryo regions or stages, however real, are not necessarily relevant for an evaluation of their developmental prospects. 相似文献
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Plasminogen activator expression in F9 teratocarcinoma embryoid bodies and their endoderm derivatives 总被引:2,自引:0,他引:2
Plasminogen activators are believed to play an important role in tissue remodeling and cell migration. During mouse embryogenesis, visceral endoderm secretes urokinase-type plasminogen activator (uPA) whereas parietal endoderm secretes tissue-type plasminogen activator (tPA). Visceral endoderm from F9 embryoid bodies can transdifferentiate into parietal endoderm under the appropriate culture conditions. We have examined at the protein and mRNA levels the type of plasminogen activator expressed in whole embryoid bodies, visceral endoderm and its parietal endoderm derivatives. Our experiments show that the visceral endoderm on F9 embryoid bodies synthesizes and secretes substantial amounts of both tPA and uPA. In contrast, the parietal endoderm derived directly from the visceral endoderm secretes dramatically increased levels of tPA and decreases production of uPA to low or below detectable levels. These data support the finding that visceral endoderm can transdifferentiate to parietal endoderm. In addition, this transition provides an excellent model for studying the molecular basis of the coincident down- and upregulation of the two plasminogen activators as well as their potential function during embryogenesis. 相似文献
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Heat shock gene expression is regulated during teratocarcinoma cell differentiation and early embryonic development 总被引:11,自引:0,他引:11
Undifferentiated teratocarcinoma stem cells do not express heat shock genes. Solid teratocarcinomas grown in vivo which contain clusters of teratocarcinoma-derived differentiated tissue do respond to heat shock. During mouse embryonic development the expression of heat shock genes is first observed with morula/blastocyst stages of mouse primplantation embryos. 相似文献
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Regulation of the cellular p53 tumor antigen in teratocarcinoma cells and their differentiated progeny. 总被引:20,自引:10,他引:10
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F9 embryonal carcinoma cells express high levels of a 53,000-molecular-weight cellular tumor antigen called p53. When F9 cell cultures are treated with retinoic acid and dibutyryl adenosine 3',5'-phosphate, they differentiate, predominantly into endoderm-like cells. This differentiation is accompanied by a marked decrease in the levels of p53. The mechanism(s) responsible for this decline in the level of p53 in differentiated cells was investigated. The results demonstrate that the high levels of p53 in F9 cells relative to their differentiated progeny were not due to alterations in the stability or turnover of this protein. Rather, the regulation during differentiation involved a marked decrease in the amount of in vitro translatable p53 mRNA detected in the differentiated cell cultures. This mechanism is unlike the one operating during the simian virus 40 infection or transformation, where the increased levels of p53 are largely due to the increased stability of the p53 protein. 相似文献
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Human embryonic stem cells have the potential to differentiate into all human cell types and therefore hold a great therapeutic promise. Differentiation into the embryonic endoderm and its derivatives is of special interest since it can provide a cure for severe widespread clinical conditions such as diabetes and hepatic failure. In this work we established a unique experimental outline that enables the study of early human endoderm development and can help improve and create new differentiation protocols. To this end we started with mesendoderm cells and separated them into early endoderm and mesoderm progenitor cells using CXCR4 and PDGFRA cell surface markers. We molecularly characterized the different lineages, and demonstrated the importance of the TGFβ pathway in definitive endoderm initiation. The endoderm progenitor cells were then purified creating an endodermal differentiation niche that is not affected by other cell populations. We followed the differentiation of these cells at different time points, and demonstrated an up regulation of genes indicative to differentiation into both foregut and hindgut. Surprisingly, upon continued culture, there was significant down regulation of the hepatic gene signature. This down regulation could be rescued with FGF2 treatment demonstrating its importance in hepatic cell maintenance. In conclusion, we suggest that isolating endoderm progenitor cells is crucial for the analysis of their fate, and enables the identification of factors involved in their differentiation and maintenance. 相似文献
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Differences in the expression of minor histocompatibility (Hm) alloantigens on two mouse embryonal carcinoma (EC) cell lines and the PYS-2 and T.D.M.-1 differentiated derivatives have been demonstrated by their ability to elicit a cytolytic T lymphocyte response. Experiments involving the use of various responder-target strain combinations and recombinant inbred mice strains have shown that: (1) there are major differences in Hm expression on EC cells compared with differentiated derivatives whose Hm expression appears more like that of adult splenocytes; (2) although both EC cell lines show reduced Hm immunogenicity compared with adult splenocytes, major differences in the expression and possible presentation of Hm between the F9 and PCC3 EC cell lines can be detected by in vivo priming and by in vitro cold competition target experiments. These observations are discussed in relation to the differences in allograft rejection patterns observed with PCC3 and F9 and to possible differences in developmental staging of these cell lines. 相似文献
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Surface proteins from five cell lines (three embryonal carcinoma cell lines (F9, PCC4 and PCC3), teratocarcinoma-derived endodermal cells (PYS) and fibroblasts (line 3/A/1-D-3 differentiated from PCC3)) were compared by two dimensional polyacrylamide gel electrophoresis after selective iodination with 125I in the presence of lactoperoxidase. The labeled proteins were solubilized either in Nonidet P40/urea/ampholyte/mercaptoethanol solution or in Nonidet P40 only. In total, about thirty major 125I-labeled surface proteins were identified by their isoelectric point and molecular weight. 14 proteins are present in all five cell types, although their quantity or accessibility for labeling differs between differentiated and undifferentiated cells. Three proteins (200, 160 and 150 kilodaltons) are present in undifferentiated cells only. Two of them (160 and 150 kilodaltons) were solubilized by Nonidet P40/urea/ampholyte/mercaptoethanol, but not by Nonidet P40. One protein (50 kilodaltons) was found in nullipotent F9 cells only. About 14–15 proteins (including fibronectin) were released by Nonidet P40/urea/ampholyte/mercaptoethanol but not by Nonidet P40. They are presumably bound to submembrane or cytoskeleton structures by non-covalent bonds. 相似文献
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Under appropriate in vitro culture conditions, mouse embryonal carcinoma can give rise to a variety of differentiated cell types, including those having the morphology of nerve cells. We report here that it is possible to evoke from such cells potential changes resembling classical neuronal action potentials, thereby indicating functional differentiation of the malignant stem cell. 相似文献
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Synthesis of hyaluronate in differentiated teratocarcinoma cells. Mechanism of chain growth 总被引:9,自引:4,他引:5
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P Prehm 《The Biochemical journal》1983,211(1):191-198
Hyaluronate could be labelled in vivo with [32P]phosphate. [32P]UDP in an alpha-glycosidic linkage constituted the reducing end of membrane-bound hyaluronate. The UDP is liberated during further chain elongation, indicating that chain growth occurs at the reducing end. [3H]Uridine could be incorporated into hyaluronate during synthesis on the isolated membraneous fraction from [3H]UDP-GlcNAc and [3H]UDP-GlcA, confirming the identification of UDP as a constituent of membrane-bound hyaluronate. These results led to a model of hyaluronate chain elongation at the reducing end by alternate addition of the chains to the substrates. Membrane-bound pyrophosphatases or 5'-nucleotidase are suggested as modulators of hyaluronate synthesis. 相似文献
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Mice obtained from blastocysts injected with malignant teratocarcinoma stem cells may comprise tumor-derived cells in their tissues. Evidence for their presence has hitherto been indirect, i.e., through detection of tissue-specific products of the tumor genotype or of strain-specific enzyme variants in tissue homogenates from healthy mice. Direct visualization and identification of the tumor-derived cells would permit their normalcy and their state of differentiation to be assessed. For this purpose, a histochemical marker is required. The marker chosen was β-galactosidase (BGS), which allows high- vs low-activity cell strains to be distinguished in situ by their differences in staining intensity. BGS has previously been employed for such visualization only in brain [Dewey, M., Gervais, A., and Mintz, B. (1976). Develop. Biol.50, 68–81] and has here been shown to be applicable to other tissues, including kidney, pancreas, and salivary gland. Two unexpected results concerning the marker itself were obtained and affected its application to histochemical comparisons: BGS activity in some tissues of some inbred strains was not concordant with that of brain, on which the existing genotypic classification is based; and some cell types within a tissue varied independently in BGS levels among strains (e.g., exocrine vs endocrine pancreas). BGS visualization clearly disclosed the presence of large numbers of fully differentiated normal cells of the teratocarcinoma strain in tissues, including the Purkinje layer of the cerebellum, the kidney tubules, and the exocrine pancreas of experimental animals. In one individual, the relevant brain region was almost entirely derived from the teratocarcinoma. Yet all tissues were indistinguishable in structure and differentiation from adult controls, and showed no malignant growth. The pattern of cell-strain distribution, which was fine-grained in the brain and patchy in the other tissues named, also resembled that of ordinary allophenic mice produced from blastomere aggregates of two strains. Thus, teratocarcinoma stem cells are here seen to undergo normal histogenesis after they are successfully incorporated into a developing host embryo. 相似文献
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Differential regulation of osteogenic marker gene expression by Wnt-3a in embryonic mesenchymal multipotential progenitor cells 总被引:6,自引:0,他引:6
Derfoul A Carlberg AL Tuan RS Hall DJ 《Differentiation; research in biological diversity》2004,72(5):209-223
The Wnt family of secreted glycoproteins plays an integral role in embryonic development and differentiation. To explore the role of Wnt's in one aspect of differentiation, namely osteogenesis, we employed a retroviral gene transfer approach to express Wnt-3a in the multipotent murine embryonic mesenchymal cell line C3H10T1/2. We found that expression of Wnt-3a in these cells had a significant, positive effect on cell growth in serum-containing medium, in that the cells grew to very high densities compared to the control cells. Additionally, apoptosis was markedly inhibited by Wnt-3a. However, when the cells were grown in serum-deficient medium, the Wnt-3a-expressing cells arrested efficiently in G1 phase, indicating that serum growth factors were needed in addition to Wnt-3a for enhanced proliferation. Wnt-3a-expressing cells exhibited high levels of alkaline phosphatase gene expression and enzymatic activity, but did not show any matrix mineralization. Unexpectedly, basal expression of bone sialoprotein, osteocalcin, and osteopontin were markedly inhibited by Wnt-3a, as were other known target genes of Wnt-3a, such as Brachyury, FGF-10, and Cdx1. When Wnt-3a-expressing cells were treated with osteogenic supplements in the presence of BMP-2, alkaline phosphatase gene expression and activity were further elevated. Additionally, BMP-2 was able to reverse the inhibitory effect of Wnt-3a on osteocalcin and osteopontin gene expression. These results indicate that while Wnt-3a represses basal expression of some osteogenic genes, this repression can be partially reversed by BMP-2. Finally, the enhanced gene expression of alkaline phosphatase induced by Wnt-3a could be effectively suppressed by the combined action of dexamethasone and 1,25-dihydroxyvitamin D(3). These data show for the first time that Wnt-3a has an unusual effect on multipotential embryonic cells, in that it enhances cellular proliferation and expression of alkaline phosphatase, while it represses most other marker genes of osteogenic differentiation. 相似文献