Prior cryopreservation of ex vivo-expanded cord blood cells is not detrimental to engraftment as measured in the NOD-SCID mouse model.
Cytokine-mediated expansion has been proposed and successfully used to facilitate engraftment post transplantation. This study examined whether cryopreservation following expansion has a detrimental effect on the ability of cells to engraft, using the NOD-SCID mouse model. Cord blood (CB) CD34(+) cells were incubated for 7 days with stem cell factor (SCF), flt-3 ligand (FL), and megakaryocyte growth and development factor (MGDF). Expanded CD34(+) cells were transplanted into NOD-SCID mice either fresh or following cryopreservation and thawing. After thawing, recovery of nucleated cells was 94%, of CD34 cells was 63%, and of day-14 progenitors was 17%. The loss of day-14 progenitor cells among the thawed expanded cells did not influence the kinetics of human engraftment in the mouse. Bone marrow (BM) of mice transplanted with thawed expanded CD34(+) cells (14 +/- 3.9%) showed significantly higher levels of human engraftment than mice transplanted with fresh expanded CD34(+) cells (1.5 +/- 0.5%, p = 0.0064). Thawed expanded CD34(+) cells had significantly higher SCID Engrafting Potential (SEP) than freshly expanded CD34(+) cells (p < 0.001). Results suggest that prior cryopreservation does not prevent expanded cells engrafting in NOD-SCID mice.
Children's Cancer Institute Australia for Medical Research, Sydney Children's Hospital, Randwick, NSW, Australia. A.Rice@unsw.edu.au
A M
AM
Rice
J A
JA
Wood
C G
CG
Milross
C J
CJ
Collins
J
J
Case
R E
RE
Nordon
M R
MR
Vowels
D000818:Animals; D018952:Antigens, CD34 / Q000097:blood; D001793:Blood Preservation / Q000379:methods* / Q000592:standards; D018929:Cell Culture Techniques; D002455:Cell Division; D015925:Cryopreservation / Q000592:standards; D005260:Female; D005312:Fetal Blood / Q000166:cytology*; D006085:Graft Survival; D018380:Hematopoietic Stem Cell Transplantation / Q000592:standards*; D006801:Humans; D017493:Leukocyte Common Antigens / Q000097:blood; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D014183:Transplantation, Heterologous
D018952:Antigens, CD34; D017493:Leukocyte Common Antigens