摘要
目的探讨人AB血浆培养外周血造血干/祖细胞(HSCs/HPCs)向成熟红细胞分化的可行性。方法将外周血HSCs/HPCs分别在添加5%胎牛血清(FBS)(FBS组)、3%FBS+2%人AB血浆(FBS+AB血浆组)和8%人AB血浆(AB血浆组)中的培养条件下诱导分化为成熟红细胞,按照1×10;/mL密度接种在24孔培养板中,2 mL/孔,观察3种培养条件下细胞生长增殖曲线和细胞形态学(细胞涂片,May-Giemsa染色,显微镜下观察细胞形态及细胞核变化)变化情况,流式细胞仪检测细胞表面红系终末分化标志分子糖化血红蛋白A(GPA)、带3蛋白(Band3)和整合素α4(α4-integrin)表达情况,以及红系晚期细胞脱核情况;比较3种培养条件对外周血HSCs/HPCs向成熟红细胞分化的影响。结果 AB血浆组、FBS组、FBS+AB血浆组的细胞生长增殖倍数分别为2 573±116 vs 2 514±246 vs 2 539±119(P>0.05);3组细胞形态学变化相似:随着培养时间延长,HSCs/HPCs从原幼红细胞向嗜碱性幼红细胞、多染幼红细胞、正染幼红细胞分化,至21 d时,几乎全部分化为脱核的红细胞;流式检测红系终末分化及脱核:血型糖蛋白A(GPA)表达及脱核率(%)分别为97.17±1.91 vs 94.95±1.61 vs 96.15±1.38、85.1±3.26 vs 86.93±5.96 vs 86.5±3.36(P>0.05).结论人AB血浆体外培养外周HSCs/HPCs向成熟红细胞分化实验达到了与胎牛血清培养相似的结果。
Objective To investigate the feasibility of differentiation of human AB plasma hematopoietic stem/progenitor cells(HSCs/HPCs) from peripheral blood into mature erythrocytes. Methods Hematopoietic stem/progenitor cells were induced to be differentiated into mature erythrocytes in the medium supplemented with 5% FBS, 3% FBS + 2% human AB plasma and 8% human AB plasma, respectively, and inoculated in 24-well culture plate at the density of 1×10;/mL. Cell proliferation and morphological changes were observed in three different groups. Flow cytometry was used to detect erythroid terminal differentiation markers, i.e. GPA, Band3 and α4(α4-integrin), and late erythroid cell enucleation in different group. The effects of different culture conditions on HSCs/HPCs differentiation into mature erythrocytes were compared.Results The cell growth and proliferation multiples of the three groups(8% human AB plasma, 5% FBS and 3% FBS+2% human AB plasma) were 2 573SymbolqB@116 vs 2 514SymbolqB@246 vs 2 539SymbolqB@119(P>0.05), respectively. The morphological changes of the three groups were similar. With the extension of culture time, the cells differentiated from proerythroblasts to basophils, polychromatic erythroblasts and positive erythroblasts, and almost all of them differentiated into erythrocytes enucleation on day 21. GPA expression and enucleation rate(%) of the three groups were 97.17SymbolqB@1.91 vs 94.95SymbolqB@1.61 vs 96.15SymbolqB@1.38, and 85.1SymbolqB@3.26 vs 86.93SymbolqB@5.96 vs 86.5SymbolqB@3.36(P>0.05), respectively. Conclusion The differentiation of HSCs/HPCs from peripheral blood plasma into mature erythrocytes from human AB was similar to that of fetal bovine serum.
作者
王姣杰
单泓
安慧娟
刘敏
别立莉
韩小改
李建斌
WANG Jiaojie;SHAN Hong;AN Huijuan;LIU Min;BIE Lili;HAN Xiaogai;LI Jianbin(Henan Red Cross Blood Center,Zhengzhou 450012,China)
出处
《中国输血杂志》
CAS
2022年第2期134-138,共5页
Chinese Journal of Blood Transfusion
基金
河南省医学科技攻关计划资助项目(201702296)。
关键词
造血干/祖细胞
体外培养
人AB血浆
胎牛血清
成熟细胞
诱导分化
hematopoietic stem/progenitor cells
in vitro culture
human AB plasma
fetal bovine serum
mature erythrocytes
induced differentiation
