Razaq Muhammad A, Taylor Stephen, Roberts David J, Carpenter Lee
Blood Research Laboratory, NHS Blood and Transplant and Nuffield Division of Clinical Laboratory Medicine, Radcliffe Department of Medicine, University of Oxford, John Radcliffe Hospital, Oxford, UK.
Computer Biology Research Group, Weatherall Institute for Molecular Medicine, Oxford, UK.
Br J Haematol. 2017 Mar;176(6):971-983. doi: 10.1111/bjh.14491. Epub 2017 Jan 6.
Human induced pluripotent stem cells (hiPSCs) are being considered for use in understanding haematopoietic disorders and as a potential source of in vitro manufactured red cells. Here, we show that hiPSCs are able to recapitulate various stages of developmental erythropoiesis. We show that primitive erythroblasts arise first, express CD31 with CD235a , embryonic globins and red cell markers, but fail to express the hallmark red cell transcripts of adult erythropoiesis. When hiPSC-derived CD45 CD235a haematopoietic progenitors are isolated on day 12 and further differentiated on OP9 stroma, they selectively express CD36 and CD235a , adult erythroid transcripts for transcription factors (e.g., BCL11A, KLF1) and fetal/adult globins (HBG1/2, HBB). Importantly, hiPSC- and cord-derived CD36 CD235a erythroblasts show a striking homology by transcriptome array profiling (only 306 transcripts with a 2Log fold change >1·5- or 2·8-fold). Phenotypic and transcriptome profiling of CD45 CD117 CD235a pro-erythroblasts and terminally differentiated erythroblasts is also provided, including evidence of a HbF (fetal) to HbA (adult) haemoglobin switch and enucleation, that mirrors their definitive erythroblast cord-derived counterparts. These findings provide a molecular roadmap of developmental erythropoiesis from hiPSC sources at several critical stages, but also helps to inform on their use for clinical applications and modelling human haematopoietic disease.
人类诱导多能干细胞（hiPSC）正被考虑用于理解造血系统疾病，并作为体外制造红细胞的潜在来源。在此，我们表明hiPSC能够重现发育性红细胞生成的各个阶段。我们发现原始成红细胞首先出现，表达CD31与CD235a、胚胎珠蛋白和红细胞标志物，但不表达成人红细胞生成的标志性红细胞转录本。当在第12天分离hiPSC来源的CD45⁻CD235a⁺造血祖细胞并在OP9基质上进一步分化时，它们选择性地表达CD36和CD235a、转录因子（如BCL11A、KLF1）以及胎儿/成人珠蛋白（HBG1/2、HBB）的成人红系转录本。重要的是，hiPSC来源和脐带血来源的CD36⁺CD235a⁺成红细胞通过转录组阵列分析显示出显著的同源性（只有306个转录本的2倍对数变化>1.5或2.8倍）。还提供了CD45⁻CD117⁻CD235a⁺早幼红细胞和终末分化成红细胞的表型和转录组分析，包括HbF（胎儿）向HbA（成人）血红蛋白转换和去核的证据，这与它们来自脐带血的确定性成红细胞对应物相似。这些发现提供了hiPSC来源在几个关键阶段发育性红细胞生成的分子路线图，也有助于为其在临床应用和人类造血疾病建模中的应用提供信息。
