Sorting and Expansion of Murine Embryonic Stem Cell Colonies Using Micropallet Arrays
Sorting and Expansion of Murine Embryonic Stem Cell Colonies Using Micropallet Arrays
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DOI：
10.1002/cyto.a.20672
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发表时间：
2009-02-01
期刊：
影响因子：
3.7
通讯作者：
Allbritton, Nancy L.
中科院分区：
文献类型：
--
作者：
Shadpour, Hamed;Sims, Christopher E.;Allbritton, Nancy L.
细胞集落的分离是大多数干细胞研究的基本任务。用于菌落选择和分离的常规技术需要大量的时间、劳动力和昂贵试剂的消耗。新的微工程技术有望通过减少所需的人力和试剂消耗来改善菌落操作。小鼠胚胎干细胞培养油阵列组成的可释放元件称为微托盘从生物相容性光致抗蚀剂创建。使用基于激光的技术释放含有未分化集落的微托盘，然后在培养物中收集细胞并扩增。微托盘阵列为维持未分化的小鼠骨骼肌细胞的培养提供了生物相容性基质。0.025%明胶的表面涂层显示出对于细胞培养和收集是最佳的。由表面粗糙的微托盘组成的阵列提供了培养和分离的进一步改进。有效地分离和收集活的斯坦细胞集落。以这种方式分选的集落显示即使在收集和在培养物中进一步扩增后仍保持未分化。定性和定量分析的分选，收集效率，和干细胞集落的释放和扩增后的细胞活力表明，micropallet阵列技术是一个很有前途的替代传统的分选方法的干细胞应用。(C)2008年国际细胞计数促进学会
Isolation of cell colonies is an essential task in most stem cell studies. Conventional techniques for colony selection and isolation require significant time, labor, and consumption of expensive reagents. New microengineered technologies hold the promise for improving colony manipulation by reducing the required manpower and reagent consumption. Murine embryonic stem cells were cultured oil arrays composed of releasable elements termed micropallets created from a biocompatible photoresist. Micropallets containing undifferentiated colonies were released using a laser-based technique followed by cell collection and expansion in Culture. The micropallet arrays provided a biocompatible Substrate for maintaining undifferentiated murine stein cells ill culture. A surface coating of 0.025% gelatin was shown to be optimal for cell culture and collection. Arrays composed of surface-roughened micropallets provided further improvements in Culture and isolation. Colonies of viable stein cells were efficiently isolated and collected. Colonies sorted in this manner were shown to remain undifferentiated even after collection and further expansion in culture. Qualitative and quantitative analyses of sorting, collection efficiency, and cell viability after release and expansion of stem cell colonies demonstrated that the micropallet array technology is a promising alternative to conventional sorting methods for stein cell applications. (C) 2008 International Society for Advancement of Cytometry