TY - JOUR
T1 - Ex vivo expansion of megakaryocyte progenitors from cryopreserved umbilical cord blood
T2 - A potential source of megakaryocytes for transplantation
AU - Pick, Marjorie
AU - Eldor, Amiram
AU - Grisaru, Dan
AU - Zander, Axel R.
AU - Shenhav, Michael
AU - Deutsch, Varda R.
N1 - Funding Information:
This work was supported by research grants from the Israel Ministry of Science (MOS), by the Cooperation Program in Cancer Research of the Deutsches Krebsforschungszentrum (DKFZ) Germany (V.R.D., A.E., and A.R.Z.) and the Israel Ministry of Health (V.D.) (The Bernard Adler Fund). The authors thank Mrs. Shoshana Bar-On for excellent technical assistance and the delivery room nurses for CB collections.
PY - 2002/9
Y1 - 2002/9
N2 - Objective. Umbilical cord blood (CB) provides an alternative source of hematopoietic progenitor cells for transplantation; however, prolonged thrombocytopenia remains a major obstacle due to the low numbers of megakaryocyte progenitor (Mk-prog) cells and their subsequent delayed engraftment. In this study, we improved techniques for enrichment, cryopreservation, and ex vivo expansion of Mk-prog cells from CB. Materials and Methods. CB mononuclear cells (MNC) were isolated and Mk-prog enriched by sedimentation on gelatin followed by centrifugation with Ficoll-Hypaque and cryopreserved. The capacity of MNC to produce Mk-prog cells, assessment of CD34+ and Mk-prog expansion in liquid culture, and analysis of the cell populations by flow cytometry were studied in cryopreserved separated CB and compared to whole CB and freshly separated samples. Results. Excellent viability of greater than 85% was maintained after cryopreservation of separated CB. The number of colony-forming Mk-prog, myeloid, and erythroid progenitor cells did not decrease with cryopreservation. Flow cytometric analysis of cryopreserved cells revealed significant removal of the residual red blood cells while maintaining complete recovery of CD34+, CD41+ (Mk), myeloid, and T and B cells compared to noncryopreserved CB cells. There was no difference in the ability of separated cryopreserved MNC CB cells to be expanded in short-term liquid cultures. Conclusions. The conditions defined here for cryopreservation of gelatin/Ficoll-Hypaque separated CB, followed by ex vivo expansion of MNC, allowed complete recovery of proliferating CD41+, CD34+, Mk-prog cells, and other hematopoietic progenitors. Mk-prog cell expansion just before the scheduled transplantation is easily applicable by this technically simple and economical procedure that requires only an aliquot of red cell cell-depleted MNC to be separated from the CB unit before cryopreservation.
AB - Objective. Umbilical cord blood (CB) provides an alternative source of hematopoietic progenitor cells for transplantation; however, prolonged thrombocytopenia remains a major obstacle due to the low numbers of megakaryocyte progenitor (Mk-prog) cells and their subsequent delayed engraftment. In this study, we improved techniques for enrichment, cryopreservation, and ex vivo expansion of Mk-prog cells from CB. Materials and Methods. CB mononuclear cells (MNC) were isolated and Mk-prog enriched by sedimentation on gelatin followed by centrifugation with Ficoll-Hypaque and cryopreserved. The capacity of MNC to produce Mk-prog cells, assessment of CD34+ and Mk-prog expansion in liquid culture, and analysis of the cell populations by flow cytometry were studied in cryopreserved separated CB and compared to whole CB and freshly separated samples. Results. Excellent viability of greater than 85% was maintained after cryopreservation of separated CB. The number of colony-forming Mk-prog, myeloid, and erythroid progenitor cells did not decrease with cryopreservation. Flow cytometric analysis of cryopreserved cells revealed significant removal of the residual red blood cells while maintaining complete recovery of CD34+, CD41+ (Mk), myeloid, and T and B cells compared to noncryopreserved CB cells. There was no difference in the ability of separated cryopreserved MNC CB cells to be expanded in short-term liquid cultures. Conclusions. The conditions defined here for cryopreservation of gelatin/Ficoll-Hypaque separated CB, followed by ex vivo expansion of MNC, allowed complete recovery of proliferating CD41+, CD34+, Mk-prog cells, and other hematopoietic progenitors. Mk-prog cell expansion just before the scheduled transplantation is easily applicable by this technically simple and economical procedure that requires only an aliquot of red cell cell-depleted MNC to be separated from the CB unit before cryopreservation.
UR - https://www.scopus.com/pages/publications/0036754605
U2 - 10.1016/S0301-472X(02)00884-6
DO - 10.1016/S0301-472X(02)00884-6
M3 - ???researchoutput.researchoutputtypes.contributiontojournal.article???
C2 - 12225800
AN - SCOPUS:0036754605
SN - 0301-472X
VL - 30
SP - 1079
EP - 1087
JO - Experimental Hematology
JF - Experimental Hematology
IS - 9
ER -