DIFFERENTIATION OF EMBRYONIC STEM CELLS: LESSONS FROM EMBRYONIC DEVELOPMENT

EMOKE PALL, ZSUZSANNA LICHNER, BABETT BONTOVICS, ELEN GOCZA

Abstract


Embryonic stem (ES) cells, the undifferentiated cells of early embryos are established as permanent lines and are characterised by their self-renewal capacity and the ability to retain their developmental capacity in vivo and in vitro. The pluripotent properties of ES cells are the basis of gene targeting technologies used to create mutant mouse strains with inactivated genes by homologous recombination. There are several methods to induce the formation of EBs. One of them the formation by aggregating ES cells in hanging drops, using gravity as an aggregation force. This method presents the advantage of obtaining well-calibrated EBs almost identical in size. We used at our experiment the mouse ES cell line KA1/11/C3/C8 with a normal karyotype, at 14th passages. Immunohistochemical examination was aimed to identify tissue-restricted proteins for the two differentiated lineages: titin as a cell-specific antigen for cardiac and skeletal muscle, betaIII-tubulin for the neuronal differentiation, cytokeratin Endo-A (TROMA) for the presence of mesenchymal progenitor cells, Oct-4 for the presence of the undifferentiated ES cells. The beating cardiac muscle clumps showed more synchronous rhythm than those seen in EBs obtained from suspension culture method, where the beating cardiac muscle clumps appeared later, had a lower frequency and were uneven. The synaptic networks of neuronal cells were best developed in EBs from suspension, compared to those observed in EBs from hanging-drop method.


Keywords


embryonic stem cells; in vitro differentiation; cardiac; neuronal

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LUCRARI STIINTIFICE ZOOTEHNIE SI BIOTEHNOLOGII (SCIENTIFIC PAPERS ANIMAL SCIENCE AND BIOTECHNOLOGIES)

ISSN print 1841 - 9364
ISSN online 2344 - 4576
ISSN-L  1841 - 9364
(former ISSN 1221-5287, E-ISSN 1841-9364)

PUBLISHER: AGROPRINT Timisoara, Romania
PAPER ACCESS: Full text articles available for free
FREQUENCY: Semiannual
PUBLICATION LANGUAGE: English
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Banat´s University of Agricultural Sciences and Veterinary Medicine "King Michael I of Romania" from Timisoara
Bioengineering Faculty of Animal Resources
300645, Timişoara, Calea Aradului 119, Romania

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