Bohmer R M, Johnson K L, Bianchi D W
Division of Genetics, Department of Pediatrics, Tufts University School of Medicine, Boston, MA, USA.
Prenat Diagn. 2001 Oct;21(10):818-23. doi: 10.1002/pd.144.
To explore potentially selective growth conditions for fetal cells in cultures from the blood of pregnant women, we investigated if fetal and adult erythroid progenitors with different hemoglobin expression programs are differentially responsive to erythropoietin (EPO). Co-cultures of clonogenic cells from 12-week fetal and adult peripheral blood were established, and the development of erythropoietic cells was monitored using flow cytometric profiles of correlated cellular contents of fetal and adult hemoglobin (HbF and HbA, respectively). Adult nucleated red cells were classified as F+A-, F+A+ or F-A+. All fetal cells were F+A-. The population of F+A- cells was flow-sorted and fetal cells were identified by fluorescence in situ hybridization (FISH) using chromosome-specific probes. Delayed EPO addition revealed that all types of erythroid cells entered the EPO-dependent phase with similar kinetics, beginning at about Day 4. The data suggest that fetal and adult erythroid stem/progenitor cells have the same initial maturation kinetics in culture independent of their hemoglobin chain expression program. Fetal and adult cells with different hemoglobin profiles also showed similar EPO dose-response curves, determined for different intervals during the first 2 weeks of culture. Thus, the kinetics of entry into the phase of EPO dependence, as well as the sensitivity to EPO at various stages of development, are essentially the same for erythropoietic progenitor cells derived from adult and early fetal blood, which rules out the possibility of using the timing or concentration of EPO for the selective growth of fetal cells from the blood of pregnant women.
为了探索孕妇血液培养物中胎儿细胞潜在的选择性生长条件，我们研究了具有不同血红蛋白表达程序的胎儿和成人红系祖细胞对促红细胞生成素（EPO）的反应是否存在差异。建立了来自12周龄胎儿和成人外周血的克隆细胞共培养体系，并使用胎儿和成人血红蛋白（分别为HbF和HbA）相关细胞含量的流式细胞术分析监测造血细胞的发育。成人有核红细胞被分类为F+A-、F+A+或F-A+。所有胎儿细胞均为F+A-。对F+A-细胞群体进行流式分选，并使用染色体特异性探针通过荧光原位杂交（FISH）鉴定胎儿细胞。延迟添加EPO显示，所有类型的红系细胞在大约第4天开始以相似的动力学进入EPO依赖期。数据表明，胎儿和成人红系干/祖细胞在培养中的初始成熟动力学相同，与其血红蛋白链表达程序无关。具有不同血红蛋白谱的胎儿和成人细胞在培养的前2周不同时间段测定的EPO剂量反应曲线也相似。因此，来自成人和早期胎儿血液的造血祖细胞进入EPO依赖期的动力学以及在发育各个阶段对EPO的敏感性基本相同，这排除了利用EPO的添加时间或浓度从孕妇血液中选择性培养胎儿细胞的可能性。
