Stem cells expressing homing receptors could be expanded from cryopreserved and unselected cord blood.
We assessed the cytokine combinations that are best for ex vivo expansion of cord blood (CB) and the increment for cell numbers of nucleated cells, as well as stem cells expressing homing receptors, by an ex vivo expansion of cryopreserved and unselected CB. Frozen leukocyte concentrates (LC) from CB were thawed and cultured at a concentration of 1 x 10(5)/mL in media supplemented with a combination of SCF (20 ng/mL)+TPO (50 ng/mL)+FL (50 ng/mL)+/-IL-6 (20 ng/mL)+/-G-CSF (20 ng/mL). After culturing for 14 days, the expansion folds of cell numbers were as follows: TNC 22.3+/-7.8 approximately 26.3+/-4.9, CFU-GM 4.7+/-5.1 approximately 11.7+/-2.6, CD34+CD38- cell 214.0+/-251.9 approximately 464.1+/-566.1, CD34+CXCR4+ cell 4384.5+/-1664.7 approximately 7087.2+/-4669.3, CD34+VLA4+ cell 1444.3+/-1264.0 approximately 2074.9+/-1537.0, CD34+VLA5+ cell 86.2+/-50.9 approximately 113.2+/-57.1. These results revealed that the number of stem cells expressing homing receptors could be increased by an ex vivo expansion of cryopreserved and unselected CB using 3 cytokines (SCF, TPO, FL) only. Further in vivo studies regarding the engraftment after expansion of the nucleated cells, as well as the stem cells expressing homing receptors will be required.
Lee
Young-Ho
YH
Hematopoietic Stem Cell Transplantation Program, Dong-A Cancer Center, Dong-A University College of Medicine, Busan, Korea. yhlee1@daunet.donga.ac.kr.
Han
Jin-Yeong
JY
Seo
Su-Yeong
SY
Kim
Kyeong-Hee
KH
Lee
Young-Ah
YA
Lee
Young-Seok
YS
Lee
Hyung-Sik
HS
Hur
Won-Joo
WJ
Han
Hun
H
Kwon
Hyuk-Chan
HC
Kim
Jae-Seok
JS
Kim
Hyo-Jin
HJ
D036541:ADP-ribosyl Cyclase / Q000378:metabolism; D051997:ADP-ribosyl Cyclase 1; D015703:Antigens, CD / Q000378:metabolism; D018952:Antigens, CD34 / Q000378:metabolism; D015925:Cryopreservation; D005312:Fetal Blood / Q000166:cytology*; D006801:Humans; D039041:Integrin alpha4beta1 / Q000378:metabolism; D039081:Integrin alpha5beta1 / Q000378:metabolism; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D019718:Receptors, CXCR4 / Q000378:metabolism; D016201:Receptors, Lymphocyte Homing / Q000378:metabolism*; D019089:Stem Cell Factor; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism*; D013926:Thrombopoietin
D015703:Antigens, CD; D018952:Antigens, CD34; D039041:Integrin alpha4beta1; D039081:Integrin alpha5beta1; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D019718:Receptors, CXCR4; D016201:Receptors, Lymphocyte Homing; D019089:Stem Cell Factor; C084881:flt3 ligand protein; D013926:Thrombopoietin; D036541:ADP-ribosyl Cyclase; C498154:CD38 protein, human; D051997:ADP-ribosyl Cyclase 1