[In vitro differentiation into megakaryocytes and generation of platelets from CD34+ cells of umbilical cord blood].
OBJECTIVE
To induce hematopoietic progenitor/stem cells of umbilical cord blood to differentiate into mature megakaryocytes and platelets in vitro and to investigate the mechanism of production of platelets.


METHODS
The CD34+ cells were sorted from umbilical cord blood by magnetic activated cell sorting (MACS) and then cultured in vitro with optimized medium to be differentiated into mature megakaryocytes and platelets. The cultured cells and the platelet-like particles were isolated from the culture and were checked by the fluorescence-activated cell sorter (FACS), immunohistochemistry assays, light microscope,electron microscope and platelet aggregation tests.


RESULTS
The cultured megakaryocytes were detected with proplatelets and both the cultured cells and the platelet-sized particles were found to have the same structure with the normal megakaryocytes and platelets by light and electron microscope. The immunohistochemistry assays revealed the cultured cells expressed GP II b III a with a positivity of 95% which was a special antigen for platelets and megakaryocytes. Culture-derived platelet-sized particles aggregated in response to thrombin as the plasma derived-platelets did. The cultured platelets had the same positivity of CD41 as the platelets from platelet rich plasma.


CONCLUSIONS
The hematopoietic progenitor/stem cells can be induced to differentiate into purified and mature megakaryocytes and platelets. It provides a practical way to study the mechanism of platelets production.
Li
Xin
X
Department of Hematology, Third Xiangya Hospital, Central South University, Changsha 410013, China.
Chen
Fang-ping
FP
Liu
Jing
J
Wu
Xin-hua
XH
Jiang
Tie-bin
TB
Tang
Xue-yuan
XY
D018952:Antigens, CD34 / Q000378:metabolism*; D001792:Blood Platelets / Q000166:cytology; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D005312:Fetal Blood / Q000166:cytology* / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D008533:Megakaryocytes / Q000166:cytology*
D018952:Antigens, CD34