PDF(Pubmed)
Abstract：
This manuscript describes step-by-step procedures to establish and manage fresh and cryopreserved cultures of nerve-derived human Schwann cells (hSCs) at the desired scale. Adaptable protocols are provided to propagate hSC cultures through serial passaging and perform routine manipulations such as enzymatic dissociation, purification, cryogenic preservation, live-cell labeling, and gene delivery. Expanded hSCs cultures are metabolically active, proliferative, and phenotypically stable for at least three consecutive passages. Cell yields are expected to be variable as determined by the rate of growth of individual batches and the rounds of subculture. The purity, however, can be maintained high at >95% hSC regardless of passage. The cells obtained in this manner are suitable for various applications, including small drug screens, in vitro modeling of neurodevelopmental processes, and cell transplantation. One caveat of this protocol is that continued expansion of same-batch hSC populations is eventually restricted due to senescence-linked growth arrest.
摘要：
该手稿描述了逐步程序，以所需的规模建立和管理神经衍生的人雪旺细胞（hSC）的新鲜和冷冻保存的培养物。提供了可适应的方案，以通过连续传代繁殖hSC培养物，并进行常规操作，例如酶促解离，净化,低温保存,活细胞标记，和基因传递。扩增的hSC培养物具有代谢活性，增殖性,并且表型稳定至少三次连续传代。细胞产量预期是可变的,如通过单个批次的生长速率和传代培养的轮次所确定的。纯度，然而,无论传代如何，都可以在>95％hSC下保持较高。以这种方式获得的细胞适用于各种应用，包括小型药物筛选，神经发育过程的体外建模，和细胞移植。该方案的一个警告是，由于衰老相关的生长停滞，最终限制了同一批hSC种群的持续扩展。
