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本帖最后由 细胞海洋 于 2011-1-6 19:26 编辑
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3 H& \: V4 I' S7 J1. 说明：原文来源自http://www.liebertonline.com/doi/abs/10.1089/scd.2010.0517 由干细胞之家新闻小组成员不倒翁翻译（转帖请注明）: @/ L6 U G9 Q! w( w1 E |
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悬浮条件下人ES和iPS未分化状态的长时间维持
- e' z. I: g' y& Q& W传统上，未分化的多能细胞，不管是人的ES还是iPS，都是在有滋养细胞或没有滋养细胞的条件下单层平铺粘附培养的，然而，多能干细胞的应用需要将培养规模扩大。因此，我们将两个ES细胞系（Royan H5和 Royan H6）和两个iPS细胞系（hiPSC1 和 hiPSC4）在无载体的悬浮条件下的聚集体进行了延长时间培养。很多代增殖之后细胞保持着稳定的核型，成功维持着未分化的多潜能状态。而且，这些细胞已于冻存和复苏而不会丧失其增值潜能、核型的稳定性以及发育的潜能。对三个细胞系的转录组分析显示，Royan H5 和 hiPSC1的贴壁培养细胞和悬浮培养的细胞几乎一致，但跟Royan H6不一样。我们仍不能确定这种转录水平上观察到的现象是否具有生物学意义。与最近的报导相比，我们的研究提供了一个hESCs 和 hiPSCs长时间培养且不丧失其冻存/复苏能力、核型稳定性和多能性的的低花费手段。我们的结果为细胞治疗、科学研究以及生物反应器系统中的工业应中所需的hESCs 和hiPSCs的扩大培养和控制分化铺平了道路。
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- e) ^; h# s( D [9 @Long-term Maintenance of Undifferentiated Human Embryonic and Induced Pluripotent Stem Cells in Suspension3 n5 p3 K8 u: Y" G% T0 p$ r- ^
Traditionally, undifferentiated pluripotent human embryonic and induced pluripotent stem cells (hESCs and hiPSCs) have been expanded as monolayer colonies in adhesion culture, both in the presence or absence of feeder cells. However, the use of pluripotent stem cells poses the need to scale-up current culture methods. Herein we present the cultivation of two hESC lines (Royan H5 and Royan H6) and two hiPSC lines (hiPSC1 and hiPSC4) as carrier-free suspension aggregates for an extended period of time. Cells proliferated over multiple passages kept a stable karyotype which successfully maintained an undifferentiated state and pluripotency, as determined by marker expressions in addition to in vitro spontaneous and directed differentiation. Additionally, these cells can be easily frozen and thawed without losing their proliferation, karyotype stability and developmental potential. Transcriptome analysis of the three lines revealed that the adherent culture condition was nearly identical to the suspension culture in Royan H5 and hiPSC1, but not in Royan H6. It remains unclear whether this observation at the transcript level is biologically significant. In comparison with recent reports, our study presents a low-cost procedure for long-term suspension expansion of hESCs and hiPSCs with the capability of freeze/thawing, karyotype stability and pluripotency. Our results will pave the way for scaled up expansion and controlled differentiation of hESCs and hiPSCs needed for cell therapy, research and industrial applications in a bioreactor culture system.
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http://www.liebertonline.com/doi/abs/10.1089/scd.2010.0517
# A( P8 I, M% m5.注明：个人翻译，理解不准确的语句还望大家积极指出。
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- e/ o: A, g9 @* L c# s2 T# V) Q4楼原文 感谢carolshen 提供 |
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发表于 2011-1-6 16:41
发表于 2011-1-6 18:12
