The purpose of the ongoing research is to improve our current skills and knowledge instem cell is... more The purpose of the ongoing research is to improve our current skills and knowledge instem cell isolation, cultivation and differentiation from the utricular and saccularepithelia of young mice. We harvested utricles and sacculi from 7 days old NMRI mice.Utricles were ...
Célunk kiméra-előállítási technikák kidolgozása volt egér, nyúl és szarvasmarha embriókon. Az egé... more Célunk kiméra-előállítási technikák kidolgozása volt egér, nyúl és szarvasmarha embriókon. Az egér ES-sejtvonalak kiméra alkotó képességét leginkább az euploid sejtek aránya befolyásolta. FISH technikát alkalmazva kimutattuk az X- és Y-kromoszómákat euploid sejtekben, illetve hiányukat az X0 kariotípusú sejtekben. Nyúl 8-sejtes morulák egyetlen sejtjének felhasználásával négy termékeny kiméra utód, két XX/XX nőstény, egy XY/XY és egy XX/XY hím született. Az egyik nőstény és az XY/XY hím germinális kiméra volt. A nyúl ES-like sejt-kolóniákat 6-7 passzázsig fenntartottuk és kidolgoztuk a sejtjeik felhasználásához szükséges kiméra technikát. Sikerült kidolgozni a tetraploid nyúl embrió előállításához szükséges paramétereket. Az A-single spermatogóniumok sejtszuszpenzióban nem azonosíthatók és kis hányadban vannak jelen, így alkalmazásuk kiméra embriók előállítására további kutatást igényel. Hatékony in vitro szarvasmarha-embrió-tenyésztési rendszert dolgoztunk ki és sikerült létrehoznu...
Romanian journal of morphology and embryology = Revue roumaine de morphologie et embryologie, 2011
Embryonic stem cells have the ability to remain undifferentiated and proliferate in vitro while m... more Embryonic stem cells have the ability to remain undifferentiated and proliferate in vitro while maintaining the potential to differentiate into derivatives of all three embryonic germ layers. The aim of the present study was to establish mouse ES lines from blastocyst stage embryos obtained after CD1/EGFP mice superovulation. We isolated, cultured and determined the characteristics of mouse embryonic stem cells in early passages, which were first described by Evans M and Kaufman M. Therefore, we evaluated the morphological criteria for the approval of ES cells in early expansion stage. Two cell lines were isolated (CDE1 and CDE2) and analyzed. They showed similar characteristics to those reported earlier for blastocyst-derived ES cell lines.
ABSTRACT In the first part of this chapter the different types of pluripotent stem cells are desc... more ABSTRACT In the first part of this chapter the different types of pluripotent stem cells are described in general: embryonal carcinoma cells, mouse and human embryonic stem cells, germ cells, epiblast stem cells and induced pluripotent cells. The methods used for isolation of rabbit embryonic stem like cells and rabbit primordial germ cells are detailed in the second part, including the species specific factors playing role in the maintenance of pluripotency. Detection of pluripotency markers both at mRNA and protein levels is an important tool in embryonic stem cell characterization. In vitro differentiation, teratoma formation and chimera forming ability are all inevitable tools to characterize embryonic stem cells. Finally the future potential of a truly validated widely available rabbit embryonic stem cell line is highlighted.
Background / Purpose: In addition to being the principal inhibitory neurotransmitter in the CNS, ... more Background / Purpose: In addition to being the principal inhibitory neurotransmitter in the CNS, GABA acts as a trophic factor during embryonic development, modulating cell proliferation, migration and differentiation. GABA is synthesized by glutamic acid decarboxylase (GAD), which exists in four different forms, namely the adult GAD65 and GAD67 and truncated embryonic GADs (EGADs), GAD25 and GAD44, respectively encoded by two alternatively spliced mRNAs that are more prevalent in the embryo. Our previous studies have shown that the four GAD forms display highly reproducible temporal expression profiles during in vivo and in vitro neuronal differentiation, prompting the idea that they play specific roles during different stages of neuronal differentiation. Here, we studied the expression of different forms of GAD in the pluripotent mouse embryonic stem (mES) and embryonal carcinoma (EC) cells to reveal the possible specific roles of these forms of GABA in the proliferation and diffe...
Although mesenchymal stem cells (MSCs) of distinct tissue origin have a large number of similarit... more Although mesenchymal stem cells (MSCs) of distinct tissue origin have a large number of similarities and differences, it has not been determined so far whether tissue-resident MSCs are the progenies of one ancestor cell lineage or the results of parallel cell developmental events. Here we compared the expression levels of 177 genes in murine MSCs derived from adult and juvenile bone marrow and adult adipose tissue, as well as juvenile spleen, thymus, and aorta wall by quantitative real-time polymerase chain reaction and the results were partially validated at protein level. All MSC lines uniformly expressed a large set of genes including well-known mesenchymal markers, such as α-smooth muscle actin, collagen type I α-chain, GATA6, Mohawk, and vimentin. In contrast, pluripotency genes and the early mesodermal marker T-gene were not expressed. On the other hand, different MSC lines consistently expressed distinct patterns of Hox genes determining the positional identity of a given cell population. Moreover, MSCs of different origin expressed a few other transcription factors also reflecting their topological identity and so the body segment or organ to which they normally contributed in vivo: (1) thymus-derived cells specifically expressed Tbx5 and Pitx2; (2) spleen-derived MSCs were characterized with Tlx1 and Nkx2.5; (3) Pitx1 designated femoral bone marrow cells and (4) En2 appeared in aorta wall-derived MSCs. Thus, MSCs exhibited topographic identity and memory even after long-term cultivation in vitro. On the basis of these results, we suggest that postnatal MSCs isolated from different anatomical sites descend from precursor cells developing in the postsegmentation mesoderm.
Pluripotent stem cells have the capacity to divide indefinitely and to differentiate into all som... more Pluripotent stem cells have the capacity to divide indefinitely and to differentiate into all somatic cells and tissue lines. They can be genetically manipulated in vitro by knocking genes in or out, and therefore serve as an excellent tool for gene function studies and for the generation of models for some human diseases. Since 1981, when the first mouse embryonic stem cell (ESC) line was generated, many attempts have been made to generate pluripotent stem cell lines from other species. Comparative characterization of ESCs from different species would help us to understand differences and similarities in the signaling pathways involved in the maintenance of pluripotency and the initiation of differentiation, and would reveal whether the fundamental mechanism controlling self-renewal of pluripotent cells is conserved across different species. This report gives an overview of research into embryonic and induced pluripotent stem cells in the rabbit, an important nonrodent species with considerable merits as an animal model for specific diseases. A number of putative rabbit ESC and induced pluripotent stem cell lines have been described. All of them expressed stem cell-associated markers and maintained apparent pluripotency during multiple passages in vitro, but none have been convincingly proven to be fully pluripotent in vivo. Moreover, as in other domestic species, the markers currently used to characterize the putative rabbit ESCs are suboptimal because recent studies have revealed that they are not always specific to the pluripotent inner cell mass. Future validation of rabbit pluripotent stem cells would benefit greatly from a validated panel of molecular markers specific to pluripotent cells of the developing rabbit embryos. Using rabbit-specific pluripotency genes may improve the efficiency of somatic cell reprogramming for generating induced pluripotent stem cells and thereby overcome some of the challenges limiting the potential of this technology.
The purpose of the ongoing research is to improve our current skills and knowledge instem cell is... more The purpose of the ongoing research is to improve our current skills and knowledge instem cell isolation, cultivation and differentiation from the utricular and saccularepithelia of young mice. We harvested utricles and sacculi from 7 days old NMRI mice.Utricles were ...
Célunk kiméra-előállítási technikák kidolgozása volt egér, nyúl és szarvasmarha embriókon. Az egé... more Célunk kiméra-előállítási technikák kidolgozása volt egér, nyúl és szarvasmarha embriókon. Az egér ES-sejtvonalak kiméra alkotó képességét leginkább az euploid sejtek aránya befolyásolta. FISH technikát alkalmazva kimutattuk az X- és Y-kromoszómákat euploid sejtekben, illetve hiányukat az X0 kariotípusú sejtekben. Nyúl 8-sejtes morulák egyetlen sejtjének felhasználásával négy termékeny kiméra utód, két XX/XX nőstény, egy XY/XY és egy XX/XY hím született. Az egyik nőstény és az XY/XY hím germinális kiméra volt. A nyúl ES-like sejt-kolóniákat 6-7 passzázsig fenntartottuk és kidolgoztuk a sejtjeik felhasználásához szükséges kiméra technikát. Sikerült kidolgozni a tetraploid nyúl embrió előállításához szükséges paramétereket. Az A-single spermatogóniumok sejtszuszpenzióban nem azonosíthatók és kis hányadban vannak jelen, így alkalmazásuk kiméra embriók előállítására további kutatást igényel. Hatékony in vitro szarvasmarha-embrió-tenyésztési rendszert dolgoztunk ki és sikerült létrehoznu...
Romanian journal of morphology and embryology = Revue roumaine de morphologie et embryologie, 2011
Embryonic stem cells have the ability to remain undifferentiated and proliferate in vitro while m... more Embryonic stem cells have the ability to remain undifferentiated and proliferate in vitro while maintaining the potential to differentiate into derivatives of all three embryonic germ layers. The aim of the present study was to establish mouse ES lines from blastocyst stage embryos obtained after CD1/EGFP mice superovulation. We isolated, cultured and determined the characteristics of mouse embryonic stem cells in early passages, which were first described by Evans M and Kaufman M. Therefore, we evaluated the morphological criteria for the approval of ES cells in early expansion stage. Two cell lines were isolated (CDE1 and CDE2) and analyzed. They showed similar characteristics to those reported earlier for blastocyst-derived ES cell lines.
ABSTRACT In the first part of this chapter the different types of pluripotent stem cells are desc... more ABSTRACT In the first part of this chapter the different types of pluripotent stem cells are described in general: embryonal carcinoma cells, mouse and human embryonic stem cells, germ cells, epiblast stem cells and induced pluripotent cells. The methods used for isolation of rabbit embryonic stem like cells and rabbit primordial germ cells are detailed in the second part, including the species specific factors playing role in the maintenance of pluripotency. Detection of pluripotency markers both at mRNA and protein levels is an important tool in embryonic stem cell characterization. In vitro differentiation, teratoma formation and chimera forming ability are all inevitable tools to characterize embryonic stem cells. Finally the future potential of a truly validated widely available rabbit embryonic stem cell line is highlighted.
Background / Purpose: In addition to being the principal inhibitory neurotransmitter in the CNS, ... more Background / Purpose: In addition to being the principal inhibitory neurotransmitter in the CNS, GABA acts as a trophic factor during embryonic development, modulating cell proliferation, migration and differentiation. GABA is synthesized by glutamic acid decarboxylase (GAD), which exists in four different forms, namely the adult GAD65 and GAD67 and truncated embryonic GADs (EGADs), GAD25 and GAD44, respectively encoded by two alternatively spliced mRNAs that are more prevalent in the embryo. Our previous studies have shown that the four GAD forms display highly reproducible temporal expression profiles during in vivo and in vitro neuronal differentiation, prompting the idea that they play specific roles during different stages of neuronal differentiation. Here, we studied the expression of different forms of GAD in the pluripotent mouse embryonic stem (mES) and embryonal carcinoma (EC) cells to reveal the possible specific roles of these forms of GABA in the proliferation and diffe...
Although mesenchymal stem cells (MSCs) of distinct tissue origin have a large number of similarit... more Although mesenchymal stem cells (MSCs) of distinct tissue origin have a large number of similarities and differences, it has not been determined so far whether tissue-resident MSCs are the progenies of one ancestor cell lineage or the results of parallel cell developmental events. Here we compared the expression levels of 177 genes in murine MSCs derived from adult and juvenile bone marrow and adult adipose tissue, as well as juvenile spleen, thymus, and aorta wall by quantitative real-time polymerase chain reaction and the results were partially validated at protein level. All MSC lines uniformly expressed a large set of genes including well-known mesenchymal markers, such as α-smooth muscle actin, collagen type I α-chain, GATA6, Mohawk, and vimentin. In contrast, pluripotency genes and the early mesodermal marker T-gene were not expressed. On the other hand, different MSC lines consistently expressed distinct patterns of Hox genes determining the positional identity of a given cell population. Moreover, MSCs of different origin expressed a few other transcription factors also reflecting their topological identity and so the body segment or organ to which they normally contributed in vivo: (1) thymus-derived cells specifically expressed Tbx5 and Pitx2; (2) spleen-derived MSCs were characterized with Tlx1 and Nkx2.5; (3) Pitx1 designated femoral bone marrow cells and (4) En2 appeared in aorta wall-derived MSCs. Thus, MSCs exhibited topographic identity and memory even after long-term cultivation in vitro. On the basis of these results, we suggest that postnatal MSCs isolated from different anatomical sites descend from precursor cells developing in the postsegmentation mesoderm.
Pluripotent stem cells have the capacity to divide indefinitely and to differentiate into all som... more Pluripotent stem cells have the capacity to divide indefinitely and to differentiate into all somatic cells and tissue lines. They can be genetically manipulated in vitro by knocking genes in or out, and therefore serve as an excellent tool for gene function studies and for the generation of models for some human diseases. Since 1981, when the first mouse embryonic stem cell (ESC) line was generated, many attempts have been made to generate pluripotent stem cell lines from other species. Comparative characterization of ESCs from different species would help us to understand differences and similarities in the signaling pathways involved in the maintenance of pluripotency and the initiation of differentiation, and would reveal whether the fundamental mechanism controlling self-renewal of pluripotent cells is conserved across different species. This report gives an overview of research into embryonic and induced pluripotent stem cells in the rabbit, an important nonrodent species with considerable merits as an animal model for specific diseases. A number of putative rabbit ESC and induced pluripotent stem cell lines have been described. All of them expressed stem cell-associated markers and maintained apparent pluripotency during multiple passages in vitro, but none have been convincingly proven to be fully pluripotent in vivo. Moreover, as in other domestic species, the markers currently used to characterize the putative rabbit ESCs are suboptimal because recent studies have revealed that they are not always specific to the pluripotent inner cell mass. Future validation of rabbit pluripotent stem cells would benefit greatly from a validated panel of molecular markers specific to pluripotent cells of the developing rabbit embryos. Using rabbit-specific pluripotency genes may improve the efficiency of somatic cell reprogramming for generating induced pluripotent stem cells and thereby overcome some of the challenges limiting the potential of this technology.
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Papers by Elen Gocza