Methodological aspects of attempts to trans-differentiate adult stem cells into embryonic-like cells in vitro.
OBJECTIVE
The aim of this research was to set up an in vitro system to trans-differentiate haematopoietic stem cells (HSCs) into embryo-like stem cells in order to de-differentiate them. In this more naive state they should be cultivated more easily in order to augment them for consecutive differentiation and autologous transplantation for use in clinical practice.


METHODS
Using the principle of the methodology of blastocyst injection, HSCs were co-cultivated with mouse embryonic stem cells (mES) with and without cell to cell contact. After co-cultivation HSCs were analyzed by flow-cytometry using 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 only finding. This implies that no de-differentiation took place.


CONCLUSIONS
We hypothesize that the unnatural situation of a mixture of two cell types originating in different species may have led to this outcome. To achieve our goal of in vitro de-differentiation we need to use a purely human culture system without animal additives.
Uher
Petr
P
Institute of Reproductive Medicine and Endocrinology, B. Smetany 2, 30177 Pilsen, Czech Republic.
Baborova
Petra
P
Huttelova
Renata
R
Kralickova
Milena
M
Vanderzwalmen
Pierre
P
Zech
Nicolas
N
D000328:Adult; D053687:Adult Stem Cells / Q000166:cytology*; D000818:Animals; D054338:Cell Transdifferentiation; D018920:Coculture Techniques; D053595:Embryonic Stem Cells / Q000166:cytology*; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D051379:Mice