Expansion and neural differentiation of embryonic stem cells in adherent and suspension cultures
Expansion and neural differentiation of embryonic stem cells in adherent and suspension cultures
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DOI：
10.1023/a:1023462832608
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发表时间：
2003-05-01
影响因子：
2.7
通讯作者：
Cabral, JMS
中科院分区：
文献类型：
--
作者：
Abranches, E;Bekman, E;Cabral, JMS
胚胎干细胞系S25是一种遗传修饰的细胞系，它允许神经细胞的谱系选择（M.利湖，加-地Lovell-Badge，A. Smith（1998）Current Biology 8：971 - 974）。在此，分析了该细胞系的生长参数。在贴壁条件下连续传代使这些细胞能够快速生长（平均比生长速率为0.035 h（-1））并产生高活细胞密度（高于90%）。还研究了S25细胞聚集成胚状体（EBs），表明有限的细胞生长（最大细胞密度为2.7 × 10（5）个细胞/ml）和聚集体大小的高度可变性（8天后为70 - 400 μ m）。用1%（v/v）胰蛋白酶酶解EB得到最高的细胞活力（91%）和密度（1.4 × 10（4）个细胞/ml），并且由此获得的细胞能够分化成神经元。
The embryonic stem cell line, S25, is a genetically modified line that allows lineage selection of neural cells (M. Li, L. Lovell-Badge, A. Smith (1998) Current Biology 8: 971-974). Here, the growth parameters of this cell line were analysed. Serial passaging in adherent conditions enabled these cells to grow rapidly (average specific growth rates of 0.035 h(-1)) and generate high viable cell densities (above 90%). The aggregation of the S25 cells into embryoid bodies (EBs) was also studied, indicating limited cell growth (maximum cell densities of 2.7 x 10(5) cells ml(-1)) and a high variability of aggregate size (70-400 mum after 8 d). Enzymatic dissociation of EBs with 1% (v/v) trypsin gave highest cell viability (91%) and density (1.4 x 10(4) cells ml(-1)) and the cells thus obtained are able to differentiate into neurons.