Expression of hyaluronan synthase genes in umbilical cord blood stem/progenitor cells.
Scientific progress reveals an ever-expanding role of hyaluronan (HA) in diverse biological functions. It has become increasingly clear that HA might also be essential for certain functions of stem cells. CD133+ cells isolated from umbilical cord blood (UCB) seem to represent an alternative to CD34+ cells as a source of transplantable haematopoietic progenitor cells. The aim of this study was to investigate expression patterns of hyaluronan synthases (HAS) genes in freshly isolated and cultured UCB progenitor cells and to compare HAS mRNA levels to those found in non-progenitor cells. CD133+ stem cells were isolated from UCB using an immunomagnetic procedure. Investigation of HAS mRNA expression patterns in CD133+ and CD133- cells by RT-PCR was performed immediately after isolation as well as after cultivation towards myelomonocytic lineage. In addition, activation patterns of mitogen activated protein kinases (MAPK) were analyzed by Western blot experiments. mRNA for HAS1 is undetectable but HAS3 mRNA can be readily detected in freshly isolated CD133+ as well as in CD133- UCB cells. More importantly, our data demonstrate that mRNA for HAS2 can only be detected in CD133+ progenitor cells. In addition, while MAPK are slightly activated in CD133- UCB cells, no significant phosphorylation of MAPK could be observed in CD133+ cells, excluding a role of these kinases in the regulation of HAS2. HAS2 is expressed only in freshly isolated CD133+ cells and quickly diminishes during differentiation. Because of this, HAS2 gene expression might be suitable as a new marker for CD133+ UCB-derived stem cells.
Institute of Laboratory Diagnostics, Ludwig Boltzmann Institute for Cardio-Thoracic Research, Kaiser Franz Joseph Hospital, Vienna, Austria.
Branka
B
Grskovic
Christine
C
Pollaschek
Mathias M
MM
Mueller
Karl M
KM
Stuhlmeier
D000071916:AC133 Antigen; D015703:Antigens, CD / Q000276:immunology; D018952:Antigens, CD34 / Q000276:immunology; D015214:Antigens, Differentiation, Myelomonocytic / Q000276:immunology; D015153:Blotting, Western; D002454:Cell Differentiation; D019070:Cell Lineage; D005312:Fetal Blood / Q000166:cytology* / Q000201:enzymology; D015971:Gene Expression Regulation, Enzymologic; D014453:Glucuronosyltransferase / Q000235:genetics*; D006023:Glycoproteins / Q000276:immunology; D006801:Humans; D000076002:Hyaluronan Synthases; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D010455:Peptides / Q000276:immunology; D019009:Proto-Oncogene Proteins c-kit / Q000276:immunology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013234:Stem Cells / Q000201:enzymology*
D000071916:AC133 Antigen; D015703:Antigens, CD; D018952:Antigens, CD34; D015214:Antigens, Differentiation, Myelomonocytic; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; D012333:RNA, Messenger; D014453:Glucuronosyltransferase; C101754:HAS2 protein, human; C105458:HAS3 protein, human; D000076002:Hyaluronan Synthases; D019009:Proto-Oncogene Proteins c-kit; D020928:Mitogen-Activated Protein Kinases