How Can the Haematopoietic Stem Cells from the Umbilical Cord Blood be De-Differentiated in vitro? Our First Results Using the Co-Cultivation Systems

Czech version

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

Overview

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