Human Schwann Cells in vitro II. Passaging, Purification, Banking, and Labeling of Established Cultures.
Human Schwann Cells in vitro II. Passaging, Purification, Banking, and Labeling of Established Cultures.
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DOI：
10.21769/bioprotoc.4882
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发表时间：
2023-11-20
期刊：
影响因子：
0.8
通讯作者：
Monje, Paula V.
中科院分区：
文献类型：
--
作者：
Monje, Paula V.
这份手稿描述了一步一步的程序来建立和管理所需规模的神经源性人类雪旺细胞(HSCs)的新鲜和冷冻培养。提供了适用的方案，通过连续传代来繁殖HSC培养物，并执行常规操作，如酶解离、纯化、低温保存、活细胞标记和基因传递。扩增的HSCs培养具有代谢活性、增殖能力和至少连续三代的表型稳定性。细胞产量预计是不同的，这取决于单个批次的生长速度和继代培养的次数。然而，无论传代与否，纯度都能在95%的HSC下保持较高的水平。以这种方式获得的细胞适合于各种应用，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，相同批次的HSC种群的持续扩张最终受到限制。
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.
影响因子：
6.2
作者：
Monje, Paula V.;Rendon, Sayuri;Athauda, Gagani;Bates, Margaret;Wood, Patrick M.;Bunge, Mary Bartlett
通讯作者：
Bunge, Mary Bartlett