Postnatal brain neural stem and progenitor cells (NSPCs) cluster in anatomically inaccessible stem cell niches, such as the subependymal zone (SEZ). Here, we describe a method for the isolation of NSPCs from live animals, which we term “milking.” The intracerebroventricular injection of a release cocktail, containing neuraminidase, integrin-β1-blocking antibody, and fibroblast growth factor 2, induces the controlled flow of NSPCs in the cerebrospinal fluid, where they are collected via liquid biopsies. Isolated cells retain key in vivo self-renewal properties and their cell-type profile reflects the cell composition of their source area, while the function of the niche is sustained even 8 months post-milking. By changing the target area more caudally, we also isolate oligodendrocyte progenitor cells (OPCs) from the corpus callosum. This novel approach for sampling NSPCs and OPCs paves the way for performing longitudinal studies in experimental animals, for more in vivo relevant cell culture assays, and for future clinical neuro-regenerative applications.

出生后脑神经干细胞和祖细胞 (NSPC) 聚集在解剖学上难以接近的干细胞壁龛中，例如室管膜下区 (SEZ)。在这里，我们描述了一种从活体动物中分离 NSPC 的方法，我们称之为“挤奶”。脑室内注射含有神经氨酸酶、整合素-β1 阻断抗体和成纤维细胞生长因子 2 的释放混合物，诱导 NSPC 在脑脊液中的受控流动，通过液体活检收集它们。分离的细胞在体内保留关键自我更新特性及其细胞类型分布反映了其来源区域的细胞组成，而生态位的功能即使在挤奶后 8 个月也能维持。通过更尾部改变目标区域，我们还从胼胝体中分离出少突胶质祖细胞 (OPCs)。这种对 NSPC 和 OPC 进行采样的新方法为在实验动物中进行纵向研究、更多的体内相关细胞培养试验以及未来的临床神经再生应用铺平了道路。