Isolation and ex vivo expansion of CD34+ cells from cord blood using dextran sedimentation and avidin column selection.
Human umbilical cord bloods were fractionated by unit gravity sedimentation in 1% (v/v) dextran, followed by immunoaffinity selection for CD34+ stem and progenitor cells. Dextran sedimentation alone enabled recovery of more than 80% of the nucleated cells present and 90% of the CD34+ cells, as determined by flow cytometry. The addition of an immunoaffinity selection step for CD34+ cells resulted in a 134-fold enrichment for CD34+ cells, with a mean yield of 64 +/- 15%. The resultant CD34+ population contained almost half the CFU-GM activity initially present in the cord bloods and could be expanded ex vivo in liquid culture.
Heimfeld
S
S
CellPro, Inc., Bothell, WA 98021.
Kalamasz
D F
DF
Fogarty
B L
BL
Fei
R
R
Tsui
Z N
ZN
Jones
H M
HM
Berenson
R J
RJ
D015703:Antigens, CD / Q000032:analysis*; D018952:Antigens, CD34; D001360:Avidin; D001710:Biotin; D001772:Blood Cell Count; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods*; D002846:Chromatography, Affinity / Q000295:instrumentation; D003911:Dextrans; D005312:Fetal Blood / Q000166:cytology*; D005427:Flocculation; D006412:Hematopoietic Stem Cells; D006801:Humans; D007231:Infant, Newborn
D015703:Antigens, CD; D018952:Antigens, CD34; D003911:Dextrans; D001360:Avidin; D001710:Biotin