Vol 47, No 2 (2009)
Original paper
Submitted: 2011-12-19
Published online: 2009-12-10
Transcriptional activity of telomerase complex in CD34- stem cells of cord blood in dependence of preparation time.
R Stojko, A Witek, J Głogowska-Lingus, U Mazurek, G Chromy, T Dworzecki, M Bojdys-Szyndlar, K Machaj, Z Pojda
DOI: 10.2478/v10042-009-0026-y
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Folia Histochem Cytobiol 2009;47(2):265-274.
Vol 47, No 2 (2009)
ORIGINAL PAPERS
Submitted: 2011-12-19
Published online: 2009-12-10
Abstract
The aim of the study was to determine whether the expression of telomerase subunits encoding genes changes during the process of cord blood preparation. It should establish if the commonly accepted 24 hours time interval in stem cells kriopreservation procedure significantly influences their immortalization and so decreases the "quality" of cord blood stem cells. Investigation includes 69 women. Spontaneous labour was the inclusion condition. The material was collected at birth after clamping of umbilical cord by direct vasopuncture. CD34- cells were extracted from cord blood (MACS, Miltenyi Biotec; Bisley, Surrey, UK). The expression profile of telomerase activators and inhibitors encoding genes was determined using HG_U133A oligonucleotide microarray (Affymetrix). We used a real-time quantitative RT-PCR assay to quantify the telomerase TERT, hTR and TP1 subunits mRNA copy numbers in CD34- cells in 0, 6, 12 and 24 hours after cord blood collection. We observed significant decrease of numbers of copies of TERTA+B mRNA within the successive hours of observation. Significant decrease of numbers of TERTA mRNA copies was confirmed after 24 hours. However, we observed significant increase of numbers of copies of TERTB mRNA after 6 hours of observation. Similar level was maintained during another 6h. The significantly lower number of copies of TERTB mRNA was observed after 24h. We also observed significant increase of number of copies of TERT mRNA after 6 hours. Number of copies of TERT mRNA significantly decreased after another 6h, remaining, however, on a higher then initial one. The significant lower number of copies of TERT mRNA was observed 24h after delivery. The possible explanation of those results is discussed in the paper.
Abstract
The aim of the study was to determine whether the expression of telomerase subunits encoding genes changes during the process of cord blood preparation. It should establish if the commonly accepted 24 hours time interval in stem cells kriopreservation procedure significantly influences their immortalization and so decreases the "quality" of cord blood stem cells. Investigation includes 69 women. Spontaneous labour was the inclusion condition. The material was collected at birth after clamping of umbilical cord by direct vasopuncture. CD34- cells were extracted from cord blood (MACS, Miltenyi Biotec; Bisley, Surrey, UK). The expression profile of telomerase activators and inhibitors encoding genes was determined using HG_U133A oligonucleotide microarray (Affymetrix). We used a real-time quantitative RT-PCR assay to quantify the telomerase TERT, hTR and TP1 subunits mRNA copy numbers in CD34- cells in 0, 6, 12 and 24 hours after cord blood collection. We observed significant decrease of numbers of copies of TERTA+B mRNA within the successive hours of observation. Significant decrease of numbers of TERTA mRNA copies was confirmed after 24 hours. However, we observed significant increase of numbers of copies of TERTB mRNA after 6 hours of observation. Similar level was maintained during another 6h. The significantly lower number of copies of TERTB mRNA was observed after 24h. We also observed significant increase of number of copies of TERT mRNA after 6 hours. Number of copies of TERT mRNA significantly decreased after another 6h, remaining, however, on a higher then initial one. The significant lower number of copies of TERT mRNA was observed 24h after delivery. The possible explanation of those results is discussed in the paper.
Title
Transcriptional activity of telomerase complex in CD34- stem cells of cord blood in dependence of preparation time.
Journal
Folia Histochemica et Cytobiologica
Issue
Vol 47, No 2 (2009)
Article type
Original paper
Pages
265-274
Published online
2009-12-10
Page views
1927
Article views/downloads
1733
DOI
10.2478/v10042-009-0026-y
Bibliographic record
Folia Histochem Cytobiol 2009;47(2):265-274.
Authors
R Stojko
A Witek
J Głogowska-Lingus
U Mazurek
G Chromy
T Dworzecki
M Bojdys-Szyndlar
K Machaj
Z Pojda
About this article