Optimization of a Method to Isolate and Culture Adult Porcine, Rats and Mice Muller Glia in Order to Study Retinal Diseases
Optimization of a Method to Isolate and Culture Adult Porcine, Rats and Mice Muller Glia in Order to Study Retinal Diseases
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DOI：
10.3389/fncel.2020.00007
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发表时间：
2020-01-29
影响因子：
5.3
通讯作者：
Vecino, Elena
中科院分区：
文献类型：
--
作者：
Pereiro, Xandra;Ruzafa, Noelia;Vecino, Elena
穆勒细胞是视网膜中主要的神经胶质细胞，垂直延伸穿过该结构，它们发挥着对神经元至关重要的丰富支持作用。米勒神经胶质细胞行为和表型的改变经常出现在视网膜变性动物模型和患有各种视网膜疾病的患者的视网膜组织中。因此，阐明视网膜疾病发生的机制将有助于更好地理解参与此类病理变化的细胞过程。由于难以获得纯细胞群以及这些细胞在培养物中快速分化的趋势，对 Muller 细胞体外活性的研究受到阻碍。目前用于分离 Muller 胶质细胞的大多数方案都使用新生儿或胚胎组织，但在这里，我们报告了一种优化的方案，有助于从成年猪、大鼠和小鼠中可靠、直接地分离和培养 Muller 细胞。这里描述的协议提供了一种快速分离成年哺乳动物 Muller 细胞的有效方法，它代表了研究治疗靶点和测试可能对抗视网膜疾病的药物效果的可靠平台。
Muller cells are the predominant glial elements in the retina, extending vertically across this structure, and they fulfill a wealth support roles that are critical for neurons. Alterations to the behavior and phenotype of Muller glia are often seen in animal models of retinal degeneration and in retinal tissue from patients with a variety of retinal disorders. Thus, elucidating the mechanisms underlying the development of retinal diseases would help better understand the cellular processes involved in such pathological changes. Studies into Muller cell activity in vitro have been hindered by the difficulty in obtaining pure cell populations and the tendency of these cells to rapidly differentiate in culture. Most protocols currently used to isolate Muller glia use neonatal or embryonic tissue but here, we report an optimized protocol that facilitates the reliable and straightforward isolation and culture of Muller cells from adult pigs, rats and mice. The protocol described here provides an efficient method for the rapid isolation of adult mammalian Muller cells, which represents a reliable platform to study therapeutic targets and to test the effects of drugs that might combat retinal diseases.