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Methodological aspects of attempts to trans-differentiate adult stem cells into embryonic like cells in vitro

Petr Uher, Petra Baborova, Renata Huttelova, Milena Kralickova, Pierre Vanderzwalmenc, Nicolas Zech

Jazyk angličtina Země Česko

Perzistentní odkaz   https://www.medvik.cz/link/bmc10009414

Grantová podpora
NR9135 MZ0 CEP - Centrální evidence projektů

Aims: The aim of this research was to set up an in vitro system to trans-diff erentiate haematopoietic stem cells(HSCs) into embryo-like stem cells in order to de-diff erentiate them. In this more naive state they should be cultivatedmore easily in order to augment them for consecutive diff erentiation and autologous transplantation for use in clinicalpractice. Methods: Using the principle of the methodology of blastocyst injection, HSCs were co-cultivated with mouseembryonic stem cells (mES) with and without cell to cell contact. After co-cultivation HSCs were analyzed by fl owcytometryusing haematopoietic markers (CD34, CD45, CD133) and embryonic stem cell markers (SSEA-4, Tra-1-60,Tra-1-81). Results: No ES cell markers were detected on the former HSCs. A decrease in HSC marker intensity was the onlyfi nding. This implies that no de-diff erentiation took place. Conclusions: We hypothesize that the unnatural situation of a mixture of two cell types originating in diff erentspecies may have led to this outcome. To achieve our goal of in vitro de-diff erentiation we need to use a purely humanculture system without animal additives.

Citace poskytuje Crossref.org

Bibliografie atd.

Lit.: 23

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$a Aims: The aim of this research was to set up an in vitro system to trans-diff erentiate haematopoietic stem cells(HSCs) into embryo-like stem cells in order to de-diff erentiate them. In this more naive state they should be cultivatedmore easily in order to augment them for consecutive diff erentiation and autologous transplantation for use in clinicalpractice. Methods: Using the principle of the methodology of blastocyst injection, HSCs were co-cultivated with mouseembryonic stem cells (mES) with and without cell to cell contact. After co-cultivation HSCs were analyzed by fl owcytometryusing haematopoietic markers (CD34, CD45, CD133) and embryonic stem cell markers (SSEA-4, Tra-1-60,Tra-1-81). Results: No ES cell markers were detected on the former HSCs. A decrease in HSC marker intensity was the onlyfi nding. This implies that no de-diff erentiation took place. Conclusions: We hypothesize that the unnatural situation of a mixture of two cell types originating in diff erentspecies may have led to this outcome. To achieve our goal of in vitro de-diff erentiation we need to use a purely humanculture system without animal additives.
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