PDF(Pubmed)
Abstract：
The enteric nervous system (ENS) is an extensive network of neurons and glia within the wall of the gastrointestinal (GI) tract that regulates many essential GI functions. Consequently, disorders of the ENS due to developmental defects, inflammation, infection, or age-associated neurodegeneration lead to serious neurointestinal diseases. Despite the prevalence and severity of these diseases, effective treatments are lacking as they fail to directly address the underlying pathology. Neuronal stem cell therapy represents a promising approach to treating diseases of the ENS by replacing the absent or injured neurons, and an autologous source of stem cells would be optimal by obviating the need for immunosuppression. We utilized the swine model to address key questions concerning cell isolation, delivery, engraftment, and fate in a large animal relevant to human therapy. We successfully isolated neural stem cells from a segment of small intestine resected from 1-month-old swine. Enteric neuronal stem cells (ENSCs) were expanded as neurospheres that grew optimally in low-oxygen (5%) culture conditions. Enteric neuronal stem cells were labeled by lentiviral green fluorescent protein (GFP) transduction, then transplanted into the same swine from which they had been harvested. Endoscopic ultrasound was then utilized to deliver the ENSCs (10,000-30,000 neurospheres per animal) into the rectal wall. At 10 and 28 days following injection, autologously derived ENSCs were found to have engrafted within rectal wall, with neuroglial differentiation and no evidence of ectopic spreading. These findings strongly support the feasibility of autologous cell isolation and delivery using a clinically useful and minimally invasive technique, bringing us closer to first-in-human ENSC therapy for neurointestinal diseases.
摘要：
肠神经系统（ENS）是胃肠道（GI）壁内神经元和神经胶质的广泛网络，可调节许多基本的GI功能。因此,由于发育缺陷导致的ENS疾病，炎症,感染,或年龄相关的神经变性导致严重的神经肠道疾病。尽管这些疾病的患病率和严重性，缺乏有效的治疗方法，因为它们不能直接解决潜在的病理学问题。神经元干细胞疗法代表了一种有希望的方法，通过替换缺失或受损的神经元来治疗ENS疾病，通过消除对免疫抑制的需要，自体干细胞来源将是最佳的。我们利用猪模型来解决有关细胞分离的关键问题，delivery,雕刻,以及与人类治疗相关的大型动物的命运。我们成功地从1个月大的猪切除的一段小肠中分离出神经干细胞。将肠神经元干细胞(ENSC)扩增为在低氧(5%)培养条件下最佳生长的神经球。通过慢病毒绿色荧光蛋白（GFP）转导标记肠神经元干细胞，然后移植到收获它们的同一只猪中。然后利用内窥镜超声将ENSC(每只动物10,000-30,000个神经球)递送到直肠壁中。注射后10天和28天，自体来源的ENSC被发现移植在直肠壁内，神经胶质分化，没有异位扩散的证据。这些发现强烈支持使用临床有用和微创技术分离和递送自体细胞的可行性，使我们更接近首次在人类ENSC治疗神经肠道疾病。
