Stem Cell

Journal of Biological Research-Thessaloniki 16: 194 – 201, 2011
J. Biol. Res.-Thessalon. is available online at http://www.jbr.gr
Indexed in: WoS (Web of Science, ISI Thomson), SCOPUS, CAS (Chemical Abstracts Service) and DOAJ (Directory of Open Access Journals)

Mesenchymal cells isolation from Wharton’s jelly, in perspective to clinical applications
Iro KOLIAKOS 1*, Nikos TSAGIAS 2 and Vassilis KARAGIANNIS 2
1

2

Stem Cells Bank, National Research Foundation, Athens, Greece
C Gynecology Clinic, Hyppokrateion Hospital, Medical School, Aristotle University of Thessaloniki, Greece
Received: 16 September 2010

Accepted after revision: 22 November 2010

Human umbilical cord tissue, termed as Wharton’s jelly, is known to be an important source of mesenchymal cells (MSCs) with considerable therapeutic potential. A rapid enzymatic isolation method in order to cryopreserve large numbers of mesenchymal cells is described and evaluated by culture expansion, flow cytometry and post-thawing viability. The purpose of this study is to provide a rapid and safe method for collection and cryopreservation of mesenchymal cells, for current and future clinical applications. Sixty-seven umbilical cords were stripped of their blood vessels, minced, enzymatically digested and cryopreserved in appropriate media containing no animal serum. MSCs were also culture expanded before cryopreservation and after thawing and characterised for MSC surface marker expression. Post-thaw viability was measured by flow cytometry. An average of 260 × 103 viable cells per cm cord were successfully isolated from all samples. Isolated cells grew adherently with a homogeneous mesenchymal morphology and expressed high levels of CD105, CD90, CD29 and low levels of CD45. They showed a high replicative potential and retained MSC marker expression over six passages. Their viability and postthaw proliferation were recorded in high values (96.9 ± 1.5%) and the expression of MSC markers was

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