A comparison of NIH-approved human ESC lines
A comparison of NIH-approved human ESC lines
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DOI：
10.1634/stemcells.2005-0452
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发表时间：
2006-12-01
期刊：
影响因子：
5.2
通讯作者：
Blau, C. Anthony
中科院分区：
文献类型：
--
作者：
Ware, Carol B.;Nelson, Angelique M.;Blau, C. Anthony
2003年10月，NIH建立了三个校外“人类胚胎干细胞研究探索中心”。“我们的中心获得了22个NIH批准的细胞系中的15个。对管路进行了以下检测：（a）不受支原体污染;（B）在自我更新和分化期间基因表达的适当模式;（c）适应均匀培养条件的能力;（d）在克隆密度下生长的能力;（e）核型;（f）生长效率;和（g）电穿孔后稳定转染的效率。一条线携带支原体。10条生产线被转换为统一条件。对9条线路进行了充分定性。人ESC（hESC）系在生长效率（通过平板培养和倍增所花费的时间量（31-57小时）测量）、克隆效率（0.8%-9.2%）和电穿孔后的稳定转染率（相对于标准小鼠ESC系为0%-53%）方面变化显著。一个hESC系在早期传代时具有不稳定的核型。需要修改与hESC供应商的拟议材料转让协议，以提高当地研究人员对hESC细胞系的可及性。NIH批准的hESC系在培养中的行为不同。许多hESC系可以使用比其供应商推荐的条件更宽松的培养条件来维持。知识产权问题对使用NIH批准的hESC系进行研究构成了重大障碍。
In October 2003, the NIH established three extramural "Exploratory Centers for Human Embryonic Stem Cell Research." Our center acquired 15 of the 22 NIH-approved cell lines. Lines were tested for: ( a) freedom from mycoplasma contamination; (b) appropriate pattern of gene expression during self-renewal and differentiation; ( c) ability to adapt to uniform culture conditions; (d) ability to grow at clonal densities; ( e) karyotype; (f) growth efficiency; and ( g) efficiency of stable transfection following electroporation. One line harbored mycoplasma. Ten lines were converted to uniform conditions. Nine lines were fully characterized. Human ESC (hESC) lines varied markedly with respect to growth efficiency as measured by the amount of time it took to plate and double (31-57 hours), cloning efficiency (0.8%-9.2%), and stable transfection rates following electroporation (0%-53% relative to a standard mouse ESC line). One hESC line had an unstable karyotype at an early passage. Modifications of the proposed Material Transfer Agreements with hESC suppliers were required to improve accessibility to hESC lines by local researchers. The NIH-approved hESC lines vary in their behavior in culture. Many hESC lines can be maintained using culture conditions less onerous than those recommended by their suppliers. Intellectual property issues pose a significant obstacle to research using NIH-approved hESC lines.