Characterisation and transplantation of enteric nervous system progenitor cells
Characterisation and transplantation of enteric nervous system progenitor cells
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DOI：
10.1136/gut.2006.094565
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发表时间：
2007-04-01
期刊：
影响因子：
24.5
通讯作者：
Edgar, David H.
中科院分区：
文献类型：
--
作者：
Almond, Sarah;Lindley, Richard M.;Edgar, David H.
目的：肠神经系统（ENS）祖细胞被认为是治疗巨结肠病的合适细胞来源。为了取得成功，以前用于分离啮齿动物ENS祖细胞的技术需要适用于出生后的人类组织。在本文中，我们描述了一种适用于制备小鼠和人出生后ENS祖细胞的方法，并评估了它们的移植潜力。方法：从交配后11.5 d的胚胎小鼠盲肠和出生后的人肌肠丛中分离单细胞悬液。这些细胞在非贴壁条件下培养产生神经球，并将其植入胚胎小鼠后肠外植体。免疫荧光显微镜观察细胞增殖、迁移和分化。结果：从小鼠和人类组织中生成的神经球含有增殖的神经嵴来源的细胞，这些细胞可以在组织培养中扩增产生胶质细胞和神经元。当植入神经节小鼠肠道时，细胞从神经球中迁移，并分化为胶质细胞和神经元，表达ENS特征的神经元表型标记，包括一氧化氮合酶和血管活性肠多肽。结论：我们已经建立了一种从新生儿中分离和扩增ENS祖细胞的技术。这些细胞在移植到神经节肠道后具有分化为神经元和胶质细胞的能力，这是它们在治疗巨结肠病的自体移植中必要的第一步。
Aims: Enteric nervous system (ENS) progenitor cells have been postulated to be an appropriate source of cells for the treatment of Hirschsprung's disease. In order for this to be successful, the techniques previously used for the isolation of rodent ENS progenitor cells need to be adapted for postnatal human tissue. In this paper, we describe a method suitable for the preparation of both mouse and human postnatal ENS progenitor cells and assess their transplantation potential.Method: Single cell suspensions were isolated from 11.5 days post-coitum embryonic mouse caecum and postnatal human myenteric plexus. These cells were cultured under non-adherent conditions to generate neurospheres which were implanted into aganglionic embryonic mouse hindgut explants. Cell proliferation, migration and differentiation were observed using immunofluorescence microscopy.Results: Neurospheres generated from both mouse and human tissues contained proliferating neural crest-derived cells that could be expanded in tissue culture to generate both glial cells and neurons. When implanted into aganglionic murine gut, cells migrated from the neurospheres using pathways appropriate for cells derived from the neural crest, and differentiated to become glia and neurons expressing neuronal phenotypic markers characteristic of the ENS including nitric oxide synthase and vasoactive intestinal polypeptide.Conclusion: We have developed a technique for the isolation and expansion of ENS progenitor cells from human neonates. These cells have the ability to differentiate into neurons and glia when transplanted into aganglionic gut, this demonstration being a necessary first step for their autologous transplantation in the treatment of Hirschsprung's disease.