Two enrichment methods to obtain CD34+ stem cells from umbilical cord blood.
We describe the relation between umbilical cord clamping time and two different enrichment system of CD34+ stem cells from umbilical cord blood with the proliferative ability and bone marrow reconstitution of the stem cells obtained. After an obstetrician performed the cord blood collection, the purification of stem cells was performed either with a combination of monoclonal antibodies (negative selections) using the Stem Sep method, or with a positive cells selection based on their surface CD34 antigens using the Mini Macs system. An excellent recovery of haematopoietic progenitors (Burst Forming Unit Erythroids (BFUE); Colony Forming Unit Granulocytes and Macrophages (CFU-GM); and Colony Forming Unit Granulocytes, Erythroids, Monocytes and Macrophages (CFU-GME)), inversely related to the increase in clamping time, was performed with the Mini Macs system (54% of colonies, with a 90% purity). With Stem Sep method, haematopoietic progenitors recovery was 35% (with a 80% purity). By applying early clamping of umbilical cord blood we obtained a greater number of CD34+ cells and their clonogenic activity was increased with enrichment. This is a useful technique considering that the number of CD34+ stem cells usually contained from a unit of placental blood is enough for the transplant to a child, but not for an adult. Thus, using these methods, we can get a larger number of CD34+ stem cells which reduces the risk of Graft versus Host Disease also in adult patients, producing survival rates similar to the those obtained with transplantation of bone marrow from unrelated donors.
Pafumi
C
C
Microbiological and Gynaecological Science Department, University of Catania, Via G. D'Annunzio no 125, 95127 Catania, Italy. pafumi@mbox.unict.it
Zizza
G
G
Pernicone
G
G
Bandiera
S
S
Farina
M
M
Russo
A
A
Iraci
M
M
Cianci
A
A
D000911:Antibodies, Monoclonal; D018952:Antigens, CD34 / Q000032:analysis*; D001781:Blood Component Removal / Q000379:methods*; D005312:Fetal Blood / Q000166:cytology*; D006412:Hematopoietic Stem Cells / Q000276:immunology*; D006801:Humans
D000911:Antibodies, Monoclonal; D018952:Antigens, CD34