Karyotyping, immunophenotyping, and apoptosis analyses on human hematopoietic precursor cells derived from umbilical cord blood following long-term ex vivo expansion.
By means of flow cytometry, CD34+/CD38- hematopoietic stem cells (HSC) were collected from umbilical cord blood (UCB) of 10 healthy women at the time of delivery and cultivated in stem-cell culture media supplemented with cell growth stimulating factors (IL-3, IL-6, GM-CSF, EPO, IGF-1, and SCF) for long periods. Apoptotic status, cell surface marker expression, and karyotypes of the cultured UCB-derived CD34+/CD38- stem-cells were investigated by flow cytometry and GTG-banding methods. The UCB-derived CD34+/CD38- stem-cells were able to divide and proliferate in vitro for at least 6 months. They did not show significantly increased apoptosis following ex vivo expansion for 20 and 32 days, respectively, in 2 cases and retained the same cell surface marker expression pattern (i.e., CD34+ and CD38-) in the majority of the cells of 2 cases following 20 and 37 days of incubation, respectively. In another 2 cases, chromosome analysis showed no evidence of numerical and structural abnormalities in the CD34+/CD38- stem-cells obtained after 20 and 43 days in culture, respectively. Our findings indicated that UCB-derived CD34+/CD38- stemcells are able to maintain their basic biologic and genetic characteristics after dividing and proliferating in vitro for a long period of time. UCB-derived HSC following ex vivo expansion can serve as a reliable resource for hematopoietic precursor cells transplantation.
Tian
Hong
H
Department of Cell Biology and Medical Genetics, Tongji Medical College of Huazhong, University of Science and Technology, Wuhan, P.R. China.
Huang
Shiang
S
Gong
Feili
F
Tian
Lei
L
Chen
Zhong
Z
D036541:ADP-ribosyl Cyclase / Q000032:analysis; D051997:ADP-ribosyl Cyclase 1; D015703:Antigens, CD / Q000032:analysis; D018952:Antigens, CD34 / Q000032:analysis; D017209:Apoptosis; D002478:Cells, Cultured; D002869:Chromosome Aberrations; D005260:Female; D005312:Fetal Blood / Q000166:cytology*; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000276:immunology* / Q000648:ultrastructure; D006801:Humans; D016130:Immunophenotyping; D007621:Karyotyping; D008562:Membrane Glycoproteins
D015703:Antigens, CD; D018952:Antigens, CD34; D008562:Membrane Glycoproteins; D036541:ADP-ribosyl Cyclase; C498154:CD38 protein, human; D051997:ADP-ribosyl Cyclase 1