title: [How can the haematopoietic stem cells from the umbilical cord blood be de-differentiated in vitro? Our first results using the co-cultivation systems].
abstract: 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.
authors.affiliation: Institut reprodukcní medicíny a endokrinologie, Plzen.
authors.forename: P
authors.initials: P
authors.lastname: Uher
authors.forename: R
authors.initials: R
authors.lastname: Hüttelová
authors.forename: M
authors.initials: M
authors.lastname: Králícková
authors.forename: Z
authors.initials: Z
authors.lastname: Novotný
authors.forename: Z
authors.initials: Z
authors.lastname: Rokyta
authors.forename: P
authors.initials: P
authors.lastname: Vanderzwalmen
authors.forename: N
authors.initials: N
authors.lastname: Zech
mesh_terms: 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