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 is...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.展开更多
We report a functionalisation strategy which is able to generate Ricinus communis agglutinin 1 (RCA 120) modified PMMA microfluidic device for binding and culturing living cells. The functionalisation is achieved by...We report a functionalisation strategy which is able to generate Ricinus communis agglutinin 1 (RCA 120) modified PMMA microfluidic device for binding and culturing living cells. The functionalisation is achieved by standard aminealdehyde (Schiff base) reaction through the crosslinker, glutaraldehyde. To prove the ability of the RCA 120 modified PMMA surface, the PC 12 cell line (rat pheochromocytoma ceils) has been captured and cultured by the microfluidic device. In the presence of tunicamycin, the dose/timedependence on decreasing of binding affinity of RCA 120 modified device with PC 12 cell is also observed. The experimental results demonstrate that the lectin-functionalized microfluidic device can be employed as efficient cell culturing platform, and has a great promise of being used as a powerful tool for monitoring the interaction of drug with living cell.展开更多
In this paper, we simply prove, in the framework of Liao, the hyperbolicity of C 1 -star invariant sets of C 1 -class differential systems on a closed manifold of dimension≤ 4, without using the C 1 -connecting lemma...In this paper, we simply prove, in the framework of Liao, the hyperbolicity of C 1 -star invariant sets of C 1 -class differential systems on a closed manifold of dimension≤ 4, without using the C 1 -connecting lemma and even the ergodic closing lemma.展开更多
Cell-cell interaction and cell metabolic analysis provide new opportunities for better understanding of critical biochemical processes. Advanced microfluidic technologies enable to create more realistic in vitro micro...Cell-cell interaction and cell metabolic analysis provide new opportunities for better understanding of critical biochemical processes. Advanced microfluidic technologies enable to create more realistic in vitro microenvironment by spatial and temporal control of cell growth and co-culture. In this work, we design a microfluidic device to achieve the co-culture of PC12 cells and 293 cells, and study in vitro cell-cell interaction via cell metabolic analysis by mass spectrometry. The membraneintegrated microfluidic device was firstly used for cell co-culture, and the cellular metabolite was further investigated by mass spectrometer(MS). Our results showed that the differentiation of PC12 cells could be successfully induced by m NGF and also greatly influenced by the microchannel treatment of fetal bovine serum(FBS) solution. The identification of cell morphology, microtubule-associated protein 2(MAP-2) expression and viability of differentiated PC12 cells were conducted before 293 cells being introduced into the top microfluidic channels and stimulated to secrete cell metabolism products. The developed microfluidic device is a potentially useful tool for high throughput of cell-cell interaction study.展开更多
基金supported by the National Natural Science Foundation of China (20733001, 20890020, 90913011, 20905004)the Ministry of Science and Technology of China (2011CB809106)+1 种基金the Ministry of Education of Chinathe Fok Ying Tung Education Foundation
文摘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.
基金supported by National Basic Research Program of China (2011CB935800)the Jilin Provincial Science and Technology Department (20100701)
文摘We report a functionalisation strategy which is able to generate Ricinus communis agglutinin 1 (RCA 120) modified PMMA microfluidic device for binding and culturing living cells. The functionalisation is achieved by standard aminealdehyde (Schiff base) reaction through the crosslinker, glutaraldehyde. To prove the ability of the RCA 120 modified PMMA surface, the PC 12 cell line (rat pheochromocytoma ceils) has been captured and cultured by the microfluidic device. In the presence of tunicamycin, the dose/timedependence on decreasing of binding affinity of RCA 120 modified device with PC 12 cell is also observed. The experimental results demonstrate that the lectin-functionalized microfluidic device can be employed as efficient cell culturing platform, and has a great promise of being used as a powerful tool for monitoring the interaction of drug with living cell.
基金supported by National Natural Science Foundation of China (Grant No. 10671088)National Basic Research Program of China (973 Project) (Grant No. 2006CB805903)
文摘In this paper, we simply prove, in the framework of Liao, the hyperbolicity of C 1 -star invariant sets of C 1 -class differential systems on a closed manifold of dimension≤ 4, without using the C 1 -connecting lemma and even the ergodic closing lemma.
基金supported by the National Natural Science Foundation of China(8137337321227006+1 种基金91213305)China Equipment and Education Resources System(CERS-1-75)
文摘Cell-cell interaction and cell metabolic analysis provide new opportunities for better understanding of critical biochemical processes. Advanced microfluidic technologies enable to create more realistic in vitro microenvironment by spatial and temporal control of cell growth and co-culture. In this work, we design a microfluidic device to achieve the co-culture of PC12 cells and 293 cells, and study in vitro cell-cell interaction via cell metabolic analysis by mass spectrometry. The membraneintegrated microfluidic device was firstly used for cell co-culture, and the cellular metabolite was further investigated by mass spectrometer(MS). Our results showed that the differentiation of PC12 cells could be successfully induced by m NGF and also greatly influenced by the microchannel treatment of fetal bovine serum(FBS) solution. The identification of cell morphology, microtubule-associated protein 2(MAP-2) expression and viability of differentiated PC12 cells were conducted before 293 cells being introduced into the top microfluidic channels and stimulated to secrete cell metabolism products. The developed microfluidic device is a potentially useful tool for high throughput of cell-cell interaction study.