Efficient stem cell isolation from under vacuum preserved tissue samples.
Different approaches for the isolation of stem/progenitor cells have been reported, including stem cell selection in stringent culture conditions. We evaluated the possibility of isolating human progenitor cells from surgical specimens preserved by under vacuum sealing and cooling, a clinical practice approached by several hospitals as alternative to formalin. Renal tissue samples (n = 20) maintained under vacuum from 6 to 48 h at 4°C were used to isolate human renal CD133(+) progenitor cells. We obtained CD133(+) progenitors from unsorted cells derived from disaggregated tissues from each sample. Phenotypic characterization as well as in vitro and in vivo differentiation of the obtained CD133(+) lines showed results comparable with sorted CD133(+) cells obtained from fresh tissue. These results indicate that the process of sealing under vacuum and cooling appears as a suitable tissue treatment to isolate hypoxia resistant cells, such as human stem/progenitor cells, and that this procedure can be exploited to render the extraction of stem cells from human samples more practical and feasible.
Department of Internal Medicine, Center for Molecular Biotechnology, Torino, Italy.
Aldo
A
Moggio
Giuseppe
G
D'Armento
Benedetta
B
Bussolati
D000071916:AC133 Antigen; D015703:Antigens, CD / Q000378:metabolism*; D061249:Batch Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D002478:Cells, Cultured; D006023:Glycoproteins / Q000378:metabolism*; D006801:Humans; D010455:Peptides / Q000378:metabolism*; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism*; D046509:Tissue Culture Techniques / Q000379:methods*; D014021:Tissue Preservation / Q000379:methods*; D014618:Vacuum
D000071916:AC133 Antigen; D015703:Antigens, CD; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides