[Formation of platelets from cord blood CD34+ cells-derived megakaryocytes induced by S-nitrosoglutathione].
To investigate the effect of S-nitrosoglutathione (GSNO), a nitric oxide donor, on platelet production from megakaryocytes differentiated from cord blood CD34(+) cells in vitro, the CD34 (+) cells from eight fresh umbilical cord blood samples by a high-gradient magnetic cell sorting (MACS) system were cultured in serum-free medium for 14 d with thrombopoietin (TPO) 50 ng/ml, IL-3 10 ng/ml, stem cell factor (SCF) 50 ng/ml and rHuGM-CSF 20 ng/ml. Then, CD61 (+) cells were purified by MACS system from these CD34 (+) cells, and were cultured in serum-free medium supplemented with TPO 50 ng/ml, IL-3 10 ng/ml and SCF 50 ng/ml in the presence (treatment group) and absence (control group) of GSNO for 30 min or 2 h. Platelet-sized particles were counted by flow cytometry; megakaryocyte structure was detected by scanning electron microscope. Aggregation of the thrombin-induced platelet particle was observed under inversion microscope. cGMP was assessed by commercial ELISA kit. The results showed that, compared with the control group, the number of platelet-sized particles significantly increased (P<0.05) in the treatment group, in which megakaryocytes presented significant pseudopod formation and extensive membrane blebbing. The platelet particle aggregation could be observed under microscope after thrombin induction. cGMP activity was significantly increased after treatment with GSNO (P<0.05). These results propose that GSNO can facilitate platelet production from megakaryocyte, and it may be partly through cGMP pathway.
Dai
Bing
B
Blood Center of Zhejiang Province, Key Laboratory of Blood Safety Research, Ministry of Health, Hangzhou 310006, China.
He
Ji
J
Chen
Shu
S
Liu
Jin-Hui
JH
Qin
Fei
F
Zhu
Fa-Ming
FM
Yan
Li-Xing
LX
D018952:Antigens, CD34 / Q000032:analysis*; D001792:Blood Platelets / Q000166:cytology*; D002454:Cell Differentiation / Q000187:drug effects*; D006152:Cyclic GMP / Q000097:blood; D005260:Female; D005312:Fetal Blood / Q000166:cytology*; D006410:Hematopoiesis / Q000187:drug effects*; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D008533:Megakaryocytes / Q000166:cytology*; D009569:Nitric Oxide / Q000502:physiology; D010974:Platelet Aggregation / Q000187:drug effects; D011247:Pregnancy; D026422:S-Nitrosoglutathione / Q000494:pharmacology*
D018952:Antigens, CD34; D009569:Nitric Oxide; D026422:S-Nitrosoglutathione; D006152:Cyclic GMP