[How can the haematopoietic stem cells from the umbilical cord blood be de-differentiated in vitro? Our first results using the co-cultivation systems].
OBJECTIVE
Aim of this study was to de-differentiate the haematopoietic stem cells (HSCs) that originated from the umbilical cord blood. One of the ways to do it is to use a co-cultivation system.


METHODS
Prospective experimental study.


METHODS
Laboratory study - Institute of reproductive medicine and endocrinology, Pilsen.


METHODS
HSCs were co-cultivated with mouse embryonic stem cells (mESC) with and without feeder cells. After co-cultivation HSCs were analyzed using flow-cytometry for presence of haematopoietic markers (CD34, CD45, CD133) and using immunohistochemistry for presence of embryonic stem cell markers (SSEA-4, Tra-1-60, Tra-1-81).


RESULTS
No de-differentiation was detectable in any our experiment, only the intensity of the HSC cell markers decreased.


CONCLUSIONS
We suppose that there were two major reasons for the experiment failure: there was no direct cell to cell contact and there was a mixture of cell types that originated from two different species. To reach our goal of in vitro de-differentiation we will need to change our strategy towards a pure human culture system without any animal additives and with cell to cell contact.
Uher
P
P
Institut reprodukcní medicíny a endokrinologie, Plzen.
Hüttelová
R
R
Králícková
M
M
Novotný
Z
Z
Rokyta
Z
Z
Vanderzwalmen
P
P
Zech
N
N
D000818:Animals; D054337:Cell Dedifferentiation; D018920:Coculture Techniques; D053595:Embryonic Stem Cells / Q000166:cytology; D005312:Fetal Blood / Q000166:cytology*; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D051379:Mice