Authors: P. Uher 1,2; R. Hűttelová 1; M. Králíčková 1,2; Z. Novotný 2; Z. Rokyta 2; P. Vanderzwalmen 3 Authors‘ workplace: Institut reprodukční medicíny a endokrinologie, Plzeň, vedoucí lékař MUDr. P. Uher 1; Gynekologicko-porodnická klinika LF UK a FN, Plzeň, přednosta doc. MUDr. Z. Rokyta, CSc. 2; Centre Hospitalier Inter Regional Cavell (CHIREC), Braine l`ąlleud, Brusel, Belgie. 3 Published in: Ceska Gynekol 2007; 72(4): 280-283 Category: Original Article
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.
Design: Prospective experimental study.
Setting: 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.
Conclusion: 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.
Key words: haematopoietic stem cells (HSCs), mouse embryonic stem cells (mESC), umbilical cord blood (UCB), flow cytometry (FACS), immunohistochemistry
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