PMID- 18928609
OWN - NLM
STAT- MEDLINE
DCOM- 20110712
LR  - 20161018
IS  - 1009-2137 (Print)
IS  - 1009-2137 (Linking)
VI  - 16
IP  - 5
DP  - 2008 Oct
TI  - [Ex vivo expansion of megakaryocyte progenitors from human umbilical cord blood 
      CD34(+) cells].
PG  - 1121-5
AB  - This study was purposed to investigate the biological characteristics and 
      immunogenicity changes of ex vivo expanded megakaryocyte progenitors from human 
      umbilical cord blood CD34(+) cells in order to provide experimental basis for 
      clinical application of ex vivo expanded umbilical cord blood megakaryocyte 
      progenitor cells. Mononuclear cells (MNCs) were obtained from umbilical cord 
      blood by Ficoll-Hyapaque density gradient separation. CD34(+) cells were enriched 
      by magnetic cell sorting (MACS). The selected CD34(+) cells were seeded in 
      serum-free medium stimulated with thrombopoietin (TPO, 50 ng/ml), interleukin 11 
      (IL-11, 50 ng/ml), and heparin (25 U/ml) for 14 days. The immunophenotyping 
      (CD34(+), CD41a(+), CD61(+), CD34(+) CD41a(+) and CD34(+) CD61(+) cells) of 
      amplificated products, matured megakaryocyte apoptosis, and expression of human 
      leukocyte antigen (HLA) class I and class II molecules were measured by 
      fluorescence-activated cell sorter (FACS). The number of colony-forming 
      units-megakaryocyte (CFU-Mk) was also evaluated by CFU-Mk assay. The results 
      showed that the umbilical cord blood CD34(+) mononuclear cells could be 
      effectively differentiated into megakaryocytes. The peak expression ratios of 
      CD41a(+) and CD61(+) cells were all at 14th days, while that of CD34(+) CD41(+) 
      and CD34(+) CD61(+) cells were at 7th day [(3.41 +/- 2.80)% and (1.89 +/- 1.43)%, 
      respectively]. The expansion times of large and small CFU-Mk reached peak at 7th 
      day (20.66 +/- 32.79) and 10th day (435.62 +/- 482.65), respectively. The 
      apoptotic rates of megakaryocytes at 7th, 10th, 14th day were (19.48 +/- 9.64)%, 
      (26.87 +/- 9.03)%, and (52.46 +/- 11.74)%, respectively. The apoptotic rate of 
      megakaryocytes had no significant difference in 7 and 10 days culture (p > 0.05), 
      while that significantly increased in culture for 14 day culture, compared with 
      culture for 7 and 10 days (p < 0.05, respectively). The expression of HLA class I 
      and class II molecules on megakaryocytes decreased along with the prolongation of 
      expansion time and sharply decreased in 0 to 10 days. It is concluded that the 
      cytokines of TPO, IL-11, and heparin can promote the expansion of megakaryocyte 
      progenitors from umbilical cord blood CD34(+) mononuclear cells effectively in 
      vitro. The peaked expansion times of large CFU-Mk, the peaked expression ratios 
      of CD34(+) CD41(+) and CD34(+) CD61(+) cells were all at 7th day. So the culture 
      for 7 days appeared to be the optimal duration of expanding megakaryocyte 
      progenitors.
FAU - He, Yi
AU  - He Y
AD  - Union Stem Cell & Gene Engineering Company LTD, Tianjin 300384, China.
FAU - Meng, Hen-Xing
AU  - Meng HX
FAU - Zhang, Yu-Guang
AU  - Zhang YG
FAU - Hou, Shi-Fang
AU  - Hou SF
FAU - Li, Qian
AU  - Li Q
FAU - Han, Jun-Ling
AU  - Han JL
FAU - Qiu, Lu-Gui
AU  - Qiu LG
FAU - Han, Zhong-Chao
AU  - Han ZC
LA  - chi
PT  - English Abstract
PT  - Journal Article
PL  - China
TA  - Zhongguo Shi Yan Xue Ye Xue Za Zhi
JT  - Zhongguo shi yan xue ye xue za zhi
JID - 101084424
RN  - 0 (Antigens, CD34)
SB  - IM
MH  - Antigens, CD34/immunology
MH  - Cell Differentiation
MH  - Cell Division
MH  - Cell Separation
MH  - Cells, Cultured
MH  - Fetal Blood/*cytology/immunology
MH  - Humans
MH  - Megakaryocyte Progenitor Cells/*cytology
EDAT- 2008/10/22 09:00
MHDA- 2011/07/13 06:00
CRDT- 2008/10/22 09:00
PHST- 2008/10/22 09:00 [pubmed]
PHST- 2011/07/13 06:00 [medline]
PHST- 2008/10/22 09:00 [entrez]
AID - 1009-2137(2008)05-1121-05 [pii]
PST - ppublish
SO  - Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2008 Oct;16(5):1121-5.