使用磁珠分选法纯化人诱导多能干细胞来源的神经前体细胞

Purification of human induced pluripotent stem cell-derived neural precursors using magnetic activated cell sorting.

作者信息

Rodrigues Gonçalo M C, Fernandes Tiago G, Rodrigues Carlos A V, Cabral Joaquim M S, Diogo Maria Margarida

机构信息

Department of Bioengineering and IBB - Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Avenida Rovisco Pais, 1049-001, Lisbon, Portugal.

出版信息

Methods Mol Biol. 2015;1283:137-45. doi: 10.1007/7651_2014_115.

DOI:10.1007/7651_2014_115

PMID:25537837

Abstract

Neural precursor (NP) cells derived from human induced pluripotent stem cells (hiPSCs), and their neuronal progeny, will play an important role in disease modeling, drug screening tests, central nervous system development studies, and may even become valuable for regenerative medicine treatments. Nonetheless, it is challenging to obtain homogeneous and synchronously differentiated NP populations from hiPSCs, and after neural commitment many pluripotent stem cells remain in the differentiated cultures. Here, we describe an efficient and simple protocol to differentiate hiPSC-derived NPs in 12 days, and we include a final purification stage where Tra-1-60+ pluripotent stem cells (PSCs) are removed using magnetic activated cell sorting (MACS), leaving the NP population nearly free of PSCs.

摘要

源自人诱导多能干细胞（hiPSC）的神经前体细胞（NP）及其神经元后代，将在疾病建模、药物筛选测试、中枢神经系统发育研究中发挥重要作用，甚至可能对再生医学治疗具有重要价值。尽管如此，从hiPSC中获得均匀且同步分化的NP群体具有挑战性，并且在神经定向分化后，许多多能干细胞仍保留在分化培养物中。在此，我们描述了一种在12天内分化hiPSC来源的NP的高效且简单的方案，并且我们包括一个最终纯化阶段，其中使用磁珠分选法（MACS）去除Tra-1-60+多能干细胞（PSC），使NP群体几乎不含PSC。