摘要
母体外周血中存在来源于胎儿的具有完整细胞核的红细胞,即胎儿有核红细胞(fetal nucleated red blood cells,FNRBCs).FNRBCs在整个妊娠期内持续存在于母体血液循环之中,具有全套的胎儿遗传物质、细胞器、各类生物分子以及代谢物等,包含丰富的胎儿生理信息,同时还表达多种特异性蛋白,是用于无创式产前检测/诊断的理想分析靶标.然而,孕妇外周血中存在数量巨大的红细胞(~10~9个/mL)和白细胞(~10~6个/mL),FNRBCs则十分稀少(<10^(3)个/mL).在完成母体静脉采血后,如何高效率高纯度分离富集血样中的FNRBCs、确定其胎儿源性并开展面向临床的生化分析极具挑战.本文从实施无创产前检测/诊断的重要意义出发,对母体外周血FNRBCs这一潜力巨大的无创产前检测/诊断标志物进行综述,在介绍FNRBCs来源及特征的基础上,总结FNRBCs分离富集与鉴定分析技术的发展现状,并阐述FNRBCs的临床应用、当前面临的挑战以及今后的发展方向,旨在为相关研究和临床转化提供参考.
Birth defect is the main reason that causes abortion,stillbirth and even the death or disability of babies,which brings great mental and economic burden to the family and the society.Traditional invasive prenatal diagnosis is highly risky and may cause damage to pregnancy women as well as their fetuses.Therefore,it is very urgent to develop safer and more convenient techniques that are able to real-time monitor the fetal health and diagnose the potential fetal diseases in time.Fetal nucleated red blood cells(FNRBCs)are found to circulate in maternal peripheral blood during the entire pregnancy while disappear rapidly after delivery.These cells not only contain the entire genome of the fetus,but also possess organelles,metabolites and biomolecules that are relating to the cellular compositions and behaviors.Therefore,FNRBCs can provide abundant materials and information for assessing the status of the fetus,which makes FNRBCs an excellent biomarker for non-invasive prenatal diagnosis.However,in 1 mL maternal peripheral blood,there are approximately 10~9 mature red blood cells(RBCs)and 10~6 white blood cells(WBCs),while only several to hundreds of FNRBCs can be acquired nowadays.The huge blood background and the rarity of FNRBCs make it a great challenge to isolate and enrich FNRBCs for downstream biomedical analysis.Furthermore,maternal nucleated red blood cells can also confuse with FNRBCs and thus hinder the accuracy and reliability of cell-based prenatal diagnosis.Conventionally,density gradient concentration,fluorescent activated cell sorting(FACS),magnetic activated cell sorting(MACS)or the combination among these three techniques are employed to isolate and enrich FNRBCs.Nevertheless,their isolation efficiency and purity are very limited.Based on newly-developed micromachining technologies,various strategies such as microfluidic chips and micro/nanostructured interfaces show excellent performance for capturing FNRBCs from maternal blood and are able to efficiently eliminate the interference of RBCs and WBCs.These strategies include immuno-affinity labeling methods aiming at antigen-antibody recognition based on specific membrane proteins of FNRBCs and label-free methods utilizing the unique physical properties of FNRBCs.Given the fact that current isolation and enrichment techniques still cannot avoid the contamination of WBCs and maternal nucleated red blood cells,it is essential to validate the fetal origin of the isolated nucleated red blood cells.Cell morphology and constitution,immunostaining of specific proteins,fluorescence in situ hybridization(FISH)and short tandem repeat(STR)analysis of unique genes are the most commonly used means to distinguish FNRBCs from other interfering cells.The identified cells can be applied in clinical non-invasive prenatal diagnosis of various diseases of both the pregnant women and their fetuses.On one hand,the abnormal variation of FNRBC enumeration during the pregnancy is related to the pathological status of either the mother(such as obesity,preeclampsia,gestational hypertension and diabetes)or the fetus(such as hypoxia and intrauterine asphyxia).On the other hand,by using molecule genetic techniques for analyzing the chromosomes and genes inside the nuclei of FNRBCs,various genetic disorders of the fetuses are able to be diagnosed,for example,13/18/21 trisomy and other aneuploidies,single gene diseases such as sickle-cell disease and thalassemia,and other chromosomal or genetic microdeletions or mutations.With the help of the development of novel and solid isolation technologies and biomedical analytical tools on the cellular/molecular level,it is greatly promising that FNRBCs will become the powerful biomarker for non-invasive prenatal diagnosis of fetal diseases.
作者
吴可
陈韦弘
何荣祥
冯春
蔡博
赵兴中
Ke Wu;Weihong Chen;Rongxiang He;Chun Feng;Bo Cai;Xingzhong Zhao(School of Environment and Health,Jianghan University,Wuhan 430056,China;School of Physics and Technology,Wuhan University,Wuhan 430072,China;College of Chemistry and Molecular Sciences,Wuhan University,Wuhan 430072,China;College of Photoelectric Materials and Technology,Jianghan University,Wuhan 430056,China;Maternal and Child Health Hospital of Hubei Province,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430070,China)
出处
《科学通报》
EI
CAS
CSCD
北大核心
2023年第31期4119-4134,共16页
Chinese Science Bulletin
基金
国家自然科学基金(81527801,82001644,81741019,61904057)资助。
关键词
胎儿有核红细胞
无创产前诊断
微流控芯片
三维微纳结构
fetal nucleated red blood cells
non-invasive prenatal diagnosis
microfluidic chips
three-dimensional micro/nano-structures