Departments of Hematology and Oncology, Hualien Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation, Hualien, Taiwan, Republic of China.
College of Medicine, Tzu-Chi University, Hualien, Taiwan, Republic of China.
Stem Cell Res Ther. 2021 May 6;12(1):274. doi: 10.1186/s13287-021-02343-4.
Hematopoiesis occurs in the bone marrow, producing a complete spectrum of blood cells to maintain homeostasis. In addition to light microscopy, chromosome analysis, and polymerase chain reaction, flow cytometry is a feasible and fast method for quantitatively analyzing hematological diseases. However, because sufficient specific cell markers are scarce, dyserythropoietic diseases are challenging to identify through flow cytometry.
Bone marrow samples from C57BL/B6 mice and one healthy donor were analyzed using traditional two-marker (CD71 and glycophorin A) flow cytometry analysis. After cell sorting, the gene expressions of membrane proteins in early and late erythropoiesis precursors and in nonerythroid cells were characterized using microarray analysis.
Among characterized gene candidates, aquaporin 0 (AQP0) expressed as a surface protein in early- and late-stage erythropoiesis precursors and was not expressed on nonerythroid cells. With the help of AQP0 staining, we could define up to five stages of erythropoiesis in both mouse and human bone marrow using flow cytometry. In addition, because patients with dyserythropoiesis generally exhibited a reduced population of APQ0 cells relative to healthy participants, the analysis results also suggested that the levels of APQ0 cells in early erythropoiesis serve as a novel biomarker that distinguishes normal from dysregulated erythropoiesis.
AQP0 was successfully demonstrated to be a marker of erythroid differentiation. The expression levels of AQP0 are downregulated in patients with dyserythropoiesis, indicating a critical role of AQP0 in erythropoiesis. Accordingly, the level of AQP0 in early erythroid precursor cells may serve as a reference parameter for diagnosing diseases associated with dyserythropoiesis.
造血发生在骨髓中，产生完整的血细胞谱以维持体内平衡。除了光学显微镜、染色体分析和聚合酶链反应外，流式细胞术是一种可行且快速的方法，可用于定量分析血液疾病。然而，由于缺乏足够的特异性细胞标志物，流式细胞术难以识别病态造血疾病。
使用传统的双标志物（CD71 和糖蛋白 A）流式细胞术分析 C57BL/B6 小鼠和一位健康供体的骨髓样本。在细胞分选后，使用微阵列分析来描述早期和晚期红细胞生成前体细胞以及非红细胞细胞中的膜蛋白的基因表达。
在鉴定的候选基因中，水通道蛋白 0（AQP0）作为早期和晚期红细胞生成前体细胞表面蛋白表达，而在非红细胞细胞中不表达。借助 AQP0 染色，我们可以使用流式细胞术在小鼠和人骨髓中定义多达五个红细胞生成阶段。此外，由于病态造血患者的 AQP0 细胞群体通常相对健康参与者减少，分析结果还表明早期红细胞生成中的 AQP0 细胞水平作为一种新的生物标志物，可区分正常和失调的红细胞生成。
AQP0 被成功证明是红细胞分化的标志物。病态造血患者的 AQP0 表达水平下调，表明 AQP0 在红细胞生成中具有关键作用。因此，早期红细胞前体细胞中 AQP0 的水平可能作为诊断与病态造血相关疾病的参考参数。
