The contribution of TPO-pathway on proliferation of megakaryocyte on co-cultured with human umbilical cord blood-derived stromal cells
- VernacularTitle:促血小板生成素途径在人脐血源基质细胞促进巨核细胞增殖中的作用
- Author:
Lei GAO
;
Xinghua CHEN
;
Xi ZHANG
- Publication Type:Journal Article
- Keywords:
stromal cells;
fetal blood;
bone marrow;
thrombopoietin (TPO);
megakaryocytes
- From:
Medical Journal of Chinese People's Liberation Army
1981;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of thrombopoietin (TPO) on the proliferation of megakaryocytic line-HEL cells co-cultured with human umbilical cord blood-derived stromal cells (hUCBDSCs), which is supposed to further elucidate the mechanism of TPO-mediated functions. Methods HEL cells were co-cultured with hUCBDSCs, and with human bone marrow stromal cells (hBMSCs), and suspended HEL was used as control. HEL The concentration of TPO in supernatant of hUCBDSCs and hBMSCs was detected by ELISA assay. The expression of c-Mpl membrane-bound protein of HEL cells was detected by laser scanning confocal microscopy and flow cytometry, and the expression of c-Mpl mRNA was detected by RT-PCR. Results It was revealed by ELISA assay that the concentration of TPO secreted by hUCBDSCs was higher than that by hBMSCs, even though the passage was done. The secretion peak in hUCBDSCs group appeared at the 8th day of culturing, somewhat delayed to that in hBMSCs group which the peak appeared at the 6th day. The expression of C-mpl protein displayed uniform green fluorescence on the surface of HEL cell line as determined by laser confocal microscopy, and mean fluorescence intensity detected by flow cytometry was 22.19?2.15, 29.65?0.82 and 37.43?1.69 in HEL suspended culture group, HEL/hBMSCs co-culture group and HEL/ hUCBDSCs co-culture group, respectively. The expression of C-mpl protein in HEL/ hUCBDSCs co-culture group was higher than that in the two other groups (P0.05). Conclusion TPO-pathway might be the important intermediate process for hUCBDSCs to promote megakaryocyte proliferation. The promotion as such may be realized by up-regulating the c-Mpl expression in translation or post-translational modification phase.
