Long term expansion of undifferentiated human iPS and ES cells in suspension culture using a defined medium
Long term expansion of undifferentiated human iPS and ES cells in suspension culture using a defined medium
复制标题
DOI：
10.1016/j.scr.2010.03.005
复制
发表时间：
2010-07-01
影响因子：
1.2
通讯作者：
Martin, Ulrich
中科院分区：
文献类型：
--
作者：
Olmer, Ruth;Haase, Alexandra;Martin, Ulrich
干细胞衍生物的治疗应用需要在成分确定的培养基中产生大量细胞，而这些细胞无法通过常规贴壁培养产生。我们应用了在OCT 4启动子控制下表达eGFP的人诱导多能干细胞（hiPS），以建立悬浮培养中未分化的人胚胎干细胞（hES）和hiPS细胞的扩增。已鉴定出一种成分确定的培养基，其在四天内导致细胞数量增加多达六倍。我们的培养系统基于初始单细胞解离，这对于标准化工艺接种至关重要。将HES / hiPS细胞扩增至17代。这些细胞保持稳定的核型、它们的多能性标记的表达以及它们分化成所有三个胚层的衍生物的潜力。在可扩展悬浮培养中扩增HES / hiPS细胞的能力代表了在搅拌生物反应器中实现标准化生产的关键一步。(C)2010 Elsevier B. V.保留所有权利。
Therapeutic application of stem cell derivatives requires large quantities of cells produced in defined media that cannot be produced via conventional adherent culture. We have applied human induced pluripotent stem (hiPS) cells expressing eGFP under control of the OCT4 promoter to establish the expansion of undifferentiated human embryonic stem (hES) and hiPS cells in suspension culture. A defined culture medium has been identified that results in up to six-fold increase in cell numbers within four days. Our culture system is based on initial single cell dissociation which is critical for standardized process inoculation. HES / hiPS cells were expanded for up to 17 passages. The cells maintained a stable karyotype, their expression of pluripotency markers and their potential to differentiate into derivatives of all three germ layers. The ability to expand HES / hiPS cells in a scalable suspension culture represents a critical step towards standardized production in stirred bioreactors. (C) 2010 Elsevier B.V. All rights reserved.