Institute of Clinical Anatomy and Cell Analysis, University of Tübingen, 72074 Tübingen, Germany.
Department of Pediatric Surgery, University Children's Hospital, 72076 Tübingen, Germany.
Cells. 2019 Jul 30;8(8):792. doi: 10.3390/cells8080792.
Identification and isolation of neural progenitor cells from the human enteric nervous system (ENS) is currently hampered by the lack of reliable, specific markers. Here, we define the Wnt-receptor frizzled-4 as a marker for the isolation of enteric neural progenitor cells derived from paediatric gut samples. We show that the Wnt-receptor frizzled-4 is expressed in the human colon and in -derived enterospheres. To obtain a purified culture, we carried out fluorescence-activated cell sorting (FACS) using PE-conjugated frizzled-4 antibodies. Frizzled-4 cells gave rise to neurosphere-like bodies and ultimately differentiated into neurons as revealed by BrdU-proliferation assays and immunocytochemistry, whereas in frizzled-4 cultures we did not detect any neuronal and glial cells. By using a patch-clamp approach, we also demonstrated the expression of functional sodium and potassium channels in frizzled-4 cell cultures after differentiation in vitro.
从人类肠神经系统 (ENS) 中分离和鉴定神经祖细胞目前受到缺乏可靠、特异标志物的限制。在这里，我们将 Wnt 受体卷曲蛋白-4 定义为分离源自儿科肠道样本的肠神经祖细胞的标志物。我们表明，Wnt 受体卷曲蛋白-4 在人类结肠和衍生的肠类器官中表达。为了获得纯化的培养物，我们使用 PE 缀合的卷曲蛋白-4 抗体进行了荧光激活细胞分选 (FACS)。卷曲蛋白-4 细胞产生神经球样体，并最终分化为神经元，如 BrdU 增殖测定和免疫细胞化学所揭示的，而在卷曲蛋白-4 培养物中，我们没有检测到任何神经元和神经胶质细胞。通过使用膜片钳技术，我们还证明了在体外分化后，卷曲蛋白-4 细胞培养物中表达功能性钠和钾通道。
