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《Chinese Journal of Blood Transfusion》 2019-01
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The effect of anti-CD36 monoclonal antibody on proliferation and differentiation of human CD34~+ hematopoietic stem (progenitor) cells

XIA Wenjie;MAI Mingqin;LI Si;CHEN Dawei;YE Xin;LI Hui;XU Xiuzahng;LEI Jinman;CHEN Yangkai;FU Yongshui;Blood Transfusion Institution of Guangzhou Blood Center;Guangdong Maternal and Child Health Hospital;The Sixth Hospital of Sun Yat-sen University;Guangdong Food and Drug Vocational College;  
Objective To investigate the effect of anti-CD36 monoclonal antibody on proliferation and differentiation of human CD34~+ hematopoietic stem(progenitor) cells in vitro. Methods Three healthy full-term maternal women without various obstetric complications were chosen, and cord blood 20 mL of each was taken. After density gradient centrifugation(800 g, 30 mins) of Ficoll cell separation solution(1.077 g/mL), CD34~+ hematopoietic stem(progenitor) cells were sorted by flow cytometry and cultured for 2—3 generations. MTT was used to examine the effect of anti-CD36 monoclonal antibody on the growth of hematopoietic stem(progenitor) cells. Flow cytometry analysis was used to detect the apoptosis and cell cycle of CD34~+ hematopoietic stem(progenitor) cells. The effect of anti-CD36 monoclonal antibody on the formation of CFU-E/BFU-E in hematopoietic stem(progenitor) cells was analyzed by CFU-E/BFU-E account after 10—14 days culture.Results After umbilical cord blood was isolated by Ficoll to obtain mononuclear cells, the hematopoietic stem(progenitor) cells of CD34~+ were sorted by flow cytometry, and about 0.44% of CD34~+ hematopoietic stem(progenitor) cells were isolated. Different concentrations of anti-CD36 monoclonal antibody(2,8,32,64 and 128 μg/mL, respectively) and hematopoietic stem(progenitor) cells were cultured in vitro. The OD value of CD36 monoclonal antibody group(2 μg/mL) was(0.9±0.15), lower than normal group(1.05±0.12)(P0.05); while the OD value(0.81±0.11) decreased significantly in the CD36 monoclonal group(32 μg/mL)(P0.01). There was no significant difference between the hematopoietic stem(progenitor) cells culture group and the IgG control group(P0.05). In the Annexin V flow detection, the apoptotic rate of anti-CD36 monoclonal antibody group(2 μg/mL) was 24.57±2.75, which was statistically higher than that of the normal group(10.13±1.42)(P0.05). Anti-CD36 monoclonal antibody significantly induced hematopoietic stem(progenitor) cells to undergo S phase cell reduction, G_1 phase cells increased, and G_1/S phase cell arrest occurred. The G_1/S value of the normal group, control group and the anti-CD36 monoclonal antibody culture group(2 μg/mL) was 3.95±0.25,4.36±0.55 and 7.35±0.65, respectively. The number of CFU-E/BFU-E clones in the normal group, IgG control group and the anti-CD36 monoclonal antibody culture group(2 μg/mL) was 169±12, 172±12, and 85±6, respectively; the number of colonies formed in the last group was significantly lower than the other two groups(P0.05).Conclusion Anti-CD36 monoclonal antibody can reduce the proliferation of human CD34~+ hematopoietic stem(progenitor) cells and reduce the ability of erythroid differentiation.
【Fund】: 广东省自然科学基金(2016A030313123 2016A030313124);△
【CateGory Index】: R714.5
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