An integrated microfluidic device for long-term culture of isolated single mammalian cells  

作　　者：ZHENG ChunHong CHEN Gui’E PANG YuHong HUANG YanYi 

机构地区：[1]College of Engineering,Peking University,Beijing 100871,China [2]Biodynamic Optical Imaging Center,Peking University,Beijing 100871,China

出　　处：《Science China Chemistry》2012年第4期502-507,共6页中国科学（化学英文版）

基　　金：supported by the National Natural Science Foundation of China (20733001, 20890020, 90913011, 20905004);the Ministry of Science and Technology of China (2011CB809106);the Ministry of Education of China;the Fok Ying Tung Education Foundation

摘　　要：We developed an integrated microfluidic chip for long-term culture of isolated single cells. This polydimethylsiloxane (PDMS) based device could accurately seed each single cell into different culture chambers, and isolate one chamber from each other with monolithically integrated pneumatic valves. We optimized the culture conditions, including the frequency of medium replacement and the introduction of conditioned medium, to keep the single cells alive for 4 days. We cultured a few hundred cells in a separated chamber on the same chip to continuously supply the conditioned medium into the culture chambers for single cells. This approach greatly facilitated the growth of single cells, and created a suitable microenvironment for observing cells' autonomous process in situ without the interference of other adjacent cells. This single cell colony assay is expandable to higher throughput, fitting the needs in the studies of drug screening and stem cell differentiation.We developed an integrated microfluidic chip for longterm culture of isolated single cells. This polydimethylsiloxane （PDMS） based device could accurately seed each single cell into different culture chambers, and isolate one chamber from each other with monolithically integrated pneumatic valves. We optimized the culture conditions, including the frequency of medium replacement and the introduction of conditioned medium, to keep the single cells alive for 4 days. We cultured a few hundred cells in a separated chamber on the same chip to continuously supply the conditioned medium into the culture chambers for single cells. This approach greatly facilitated the growth of single cells, and created a suitable microenvironment for observing cells＇ autonomous process in situ without the interference of other adjacent cells. This single cell colony assay is expandable to higher throughput, fitting the needs in the studies of drug screening and stem cell differentiation.

关 键 词：MICROFLUIDICS single cell culture conditioned medium 

分 类 号：Q78[生物学—分子生物学] Q954.6