Optimization of immunomagnetic separation for cord blood-derived hematopoietic stem cells.
BACKGROUND
There is a growing interest in cord blood as a source of primitive stem cells with the capacity for multilineage differentiation. Pure cell fractions are needed for the characterization and in vitro expansion of stem cells as well as for their use in preclinical research. However, enrichment of stem cells is challenging due to the lack of stem cell-specific markers and gentle protocols for the isolation of highly pure stem cell fractions. Protocols developed for the enrichment of peripheral blood-derived stem cells have been found to be suboptimal for cord blood.


RESULTS
In this study, immunomagnetic cell sorting protocols to purify CD34+, CD133+ and Lin- cells from fresh and cryopreserved cord blood were optimized. Reproducible purities of up to 97% were reached. The selected cells were highly viable having substantial colony-forming potential.


CONCLUSIONS
The optimized protocols enable rapid enrichment of highly pure hematopoietic stem cells from both fresh and cryopreserved cord blood.
Kekarainen
Tuija
T
Finnish Red Cross Blood Service, Helsinki, Finland. tuija.kekarainen@cresa.uab.es
Mannelin
Sirkka
S
Laine
Jarmo
J
Jaatinen
Taina
T
D000071916:AC133 Antigen; D015703:Antigens, CD / Q000378:metabolism; D018952:Antigens, CD34 / Q000378:metabolism; D002469:Cell Separation / Q000379:methods*; D003114:Colony-Forming Units Assay; D015925:Cryopreservation; D005312:Fetal Blood / Q000166:cytology*; D006023:Glycoproteins / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods*; D007958:Leukocyte Count; D007962:Leukocytes / Q000166:cytology; D010455:Peptides / Q000378:metabolism
D000071916:AC133 Antigen; D015703:Antigens, CD; D018952:Antigens, CD34; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides